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| <head> | | <head> |
− | <meta charset="utf-8"/> | + | <meta charset="utf-8" /> |
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| <title>Proof Of Concept | iGEM HUST-China</title> | | <title>Proof Of Concept | iGEM HUST-China</title> |
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− | rel="stylesheet"/>
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− | <body><!-- # TODO: #6 Fix table caption font--><!-- # TODO: #7 Fix citations links font size--> | + | |
− | <nav class="navbar navbar-expand-xl fixed-top"> | + | <body> |
− | <div class="container d-flex justify-content-between"><a class="navbar-brand"
| + | <!-- # TODO: #6 Fix table caption font--> |
− | href="https://2021.igem.org/Team:HUST-China"><i
| + | <!-- # TODO: #7 Fix citations links font size--> |
− | class="navbar-logo-left"></i><span>HUST-China</span></a>
| + | <nav class="navbar navbar-expand-xl fixed-top"> |
− | <button aria-controls="navbarNav" aria-expanded="false" aria-label="Toggle navigation" class="navbar-toggler"
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− | data-target="#navbarNav" data-toggle="collapse" type="button"><span class="navbar-toggler-icon"></span>
| + | href="https://2021.igem.org/Team:HUST-China"><i class="navbar-logo-left"></i><span>HUST-China</span></a> |
− | </button>
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− | <div class="collapse navbar-collapse" id="navbarNav">
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− | <li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true"
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− | class="nav-link dropdown-toggle" data-toggle="dropdown" href="#"
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− | id="navbarProjectDropdown" role="button">Project</a>
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− | <div aria-labelledby="navbarProjectDropdown" class="dropdown-menu"><a class="dropdown-item"
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− | href="https://2021.igem.org/Team:HUST-China/Description">Description</a><a
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− | class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Design">Design</a><a
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− | class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Proof_Of_Concept">Proof of
| + | <div aria-labelledby="navbarProjectDropdown" class="dropdown-menu"><a class="dropdown-item" |
− | Concept</a><a class="dropdown-item"
| + | href="https://2021.igem.org/Team:HUST-China/Description">Description</a><a |
− | href="https://2021.igem.org/Team:HUST-China/Safety">Safety</a><a
| + | class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Design">Design</a><a |
− | class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Implementation">Proposed
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− | Implementation</a><a class="dropdown-item"
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− | href="https://2021.igem.org/Team:HUST-China/Contribution">Contribution</a>
| + | Concept</a><a class="dropdown-item" |
− | </div>
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− | </li>
| + | href="https://2021.igem.org/Team:HUST-China/Implementation">Proposed |
− | <li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true"
| + | Implementation</a><a class="dropdown-item" |
− | class="nav-link dropdown-toggle" data-toggle="dropdown" href="#"
| + | href="https://2021.igem.org/Team:HUST-China/Contribution">Contribution</a> |
− | id="navbarHuman PracticesDropdown" role="button">Human Practices</a>
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− | <div aria-labelledby="navbarHuman PracticesDropdown" class="dropdown-menu"><a class="dropdown-item"
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− | href="https://2021.igem.org/Team:HUST-China/Human_Practices">Overview</a><a
| + | <li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true" |
− | class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Human_Practices_01">Part
| + | class="nav-link dropdown-toggle" data-toggle="dropdown" href="#" |
− | 1</a><a class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Human_Practices_02">Part
| + | id="navbarHuman PracticesDropdown" role="button">Human Practices</a> |
− | 2</a><a class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Collaborations">Part
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− | 3</a><a class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Education">Part 4</a>
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− | </div>
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− | </li>
| + | class="dropdown-item" |
− | <li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true"
| + | href="https://2021.igem.org/Team:HUST-China/Human_Practices_01">Part |
− | class="nav-link dropdown-toggle" data-toggle="dropdown" href="#"
| + | 1</a><a class="dropdown-item" |
− | id="navbarModelingDropdown" role="button">Modeling</a>
| + | href="https://2021.igem.org/Team:HUST-China/Human_Practices_02">Part |
− | <div aria-labelledby="navbarModelingDropdown" class="dropdown-menu"><a class="dropdown-item"
| + | 2</a><a class="dropdown-item" |
− | href="https://2021.igem.org/Team:HUST-China/Model">Overview</a><a
| + | href="https://2021.igem.org/Team:HUST-China/Collaborations">Part |
− | class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Gene_Pathway">Gene
| + | 3</a><a class="dropdown-item" |
− | Pathway</a><a class="dropdown-item"
| + | href="https://2021.igem.org/Team:HUST-China/Education">Part 4</a> |
− | href="https://2021.igem.org/Team:HUST-China/Dye_color_forecast">Dye color
| + | </div> |
− | forecast</a><a class="dropdown-item"
| + | </li> |
− | href="https://2021.igem.org/Team:HUST-China/Micronucleus_Counter">Micronucleus
| + | <li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true" |
− | Counter</a></div>
| + | class="nav-link dropdown-toggle" data-toggle="dropdown" href="#" id="navbarModelingDropdown" |
− | </li>
| + | role="button">Modeling</a> |
− | <li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true"
| + | <div aria-labelledby="navbarModelingDropdown" class="dropdown-menu"><a class="dropdown-item" |
− | class="nav-link dropdown-toggle" data-toggle="dropdown" href="#"
| + | href="https://2021.igem.org/Team:HUST-China/Model">Overview</a><a class="dropdown-item" |
− | id="navbarProductsDropdown" role="button">Products</a>
| + | href="https://2021.igem.org/Team:HUST-China/Gene_Pathway">Gene |
− | <div aria-labelledby="navbarProductsDropdown" class="dropdown-menu"><a class="dropdown-item"
| + | Pathway</a><a class="dropdown-item" |
− | href="https://2021.igem.org/Team:HUST-China/Products">Overview</a><a
| + | href="https://2021.igem.org/Team:HUST-China/Dye_color_forecast">Dye color |
− | class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Software">Software</a><a
| + | forecast</a><a class="dropdown-item" |
− | class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Hardware">Hardware</a><a
| + | href="https://2021.igem.org/Team:HUST-China/Micronucleus_Counter">Micronucleus |
− | class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Another_Area">Another
| + | Counter</a></div> |
− | Area</a></div>
| + | </li> |
− | </li>
| + | <li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true" |
− | <li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true"
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− | class="nav-link dropdown-toggle" data-toggle="dropdown" href="#"
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− | id="navbarExperimentsDropdown" role="button">Experiments</a>
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− | <div aria-labelledby="navbarExperimentsDropdown" class="dropdown-menu"><a class="dropdown-item"
| + | href="https://2021.igem.org/Team:HUST-China/Products">Overview</a><a |
− | href="https://2021.igem.org/Team:HUST-China/Experiments">Overview</a><a
| + | class="dropdown-item" |
− | class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Parts">Parts</a><a
| + | href="https://2021.igem.org/Team:HUST-China/Software">Software</a><a |
− | class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Engineering">Engineering
| + | class="dropdown-item" |
− | Success</a><a class="dropdown-item"
| + | href="https://2021.igem.org/Team:HUST-China/Hardware">Hardware</a><a |
− | href="https://2021.igem.org/Team:HUST-China/Results">Results</a><a
| + | class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Another_Area">Another |
− | class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Protocol">Protocol</a><a
| + | Area</a></div> |
− | class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Notebook">Notebook</a>
| + | </li> |
− | </div>
| + | <li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true" |
− | </li>
| + | class="nav-link dropdown-toggle" data-toggle="dropdown" href="#" |
− | <li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true"
| + | id="navbarExperimentsDropdown" role="button">Experiments</a> |
− | class="nav-link dropdown-toggle" data-toggle="dropdown" href="#"
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− | id="navbarPeopleDropdown" role="button">People</a>
| + | href="https://2021.igem.org/Team:HUST-China/Experiments">Overview</a><a |
− | <div aria-labelledby="navbarPeopleDropdown" class="dropdown-menu"><a class="dropdown-item"
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− | href="https://2021.igem.org/Team:HUST-China/Team">Team</a><a
| + | class="dropdown-item" |
− | class="dropdown-item"
| + | href="https://2021.igem.org/Team:HUST-China/Engineering">Engineering |
− | href="https://2021.igem.org/Team:HUST-China/Partnership">Partnership</a><a
| + | Success</a><a class="dropdown-item" |
− | class="dropdown-item"
| + | href="https://2021.igem.org/Team:HUST-China/Results">Results</a><a class="dropdown-item" |
− | href="https://2021.igem.org/Team:HUST-China/Attributions">Attributions</a></div>
| + | href="https://2021.igem.org/Team:HUST-China/Protocol">Protocol</a><a |
− | </li>
| + | class="dropdown-item" href="https://2021.igem.org/Team:HUST-China/Notebook">Notebook</a> |
− | <li class="nav-item dropdown"><a aria-expanded="false" aria-haspopup="true"
| + | </div> |
− | class="nav-link dropdown-toggle" data-toggle="dropdown" href="#"
| + | </li> |
− | id="navbarAchievementsDropdown" role="button">Achievements</a>
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− | <div aria-labelledby="navbarAchievementsDropdown" class="dropdown-menu"><a class="dropdown-item"
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− | href="https://2021.igem.org/Team:HUST-China/Judging">Judging</a><a
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− | class="dropdown-item"
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− | href="https://2021.igem.org/Team:HUST-China/https:/video.igem.org/w/aMwNT6eBryGs8xG5avXD51">Promotion
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− | Video</a><a class="dropdown-item"
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− | Form</a></div>
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− | </li>
| + | </li> |
− | </ul>
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− | </div>
| + | class="nav-link dropdown-toggle" data-toggle="dropdown" href="#" |
− | </div>
| + | id="navbarAchievementsDropdown" role="button">Achievements</a> |
− | </nav>
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− | <div class="navbar-extra fixed-top"></div>
| + | href="https://2021.igem.org/Team:HUST-China/Award">Award</a><a class="dropdown-item" |
− | <header class="d-flex justify-content-center align-items-center">
| + | href="https://video.igem.org/w/aMwNT6eBryGs8xG5avXD51">Promotion |
− | <div class="container"><h1 id="Top">Proof Of Concept</h1>
| + | Video</a><a class="dropdown-item" |
− | <p class="lead pl-1"></p>
| + | href="https://igem.org/2021_Judging_Form?id=3711">Judging |
− | <hr class="my-4"/>
| + | Form</a></div> |
− | </div>
| + | </li> |
− | </header>
| + | </ul> |
− | <main>
| + | |
− | <div class="container">
| + | |
− | <div class="row">
| + | |
− | <div class="sidebar col-lg-3">
| + | |
− | <div class="nav" id="contents"><h5>Contents</h5> | + | |
− | <ul></ul>
| + | |
− | </div>
| + | |
| </div> | | </div> |
− | <div class="content col-lg-9">
| + | </div> |
− | <article><h1>Introduction</h1>
| + | </nav> |
− | <p>Our project (click <a
| + | <div class="navbar-extra fixed-top"></div> |
− | href="https://2021.igem.org/Team:HUST-China/Proof_Of_Concept/Design">design</a> for detailed
| + | <header class="d-flex justify-content-center align-items-center"> |
− | information) integrated at most five reactions within a system. Each reaction in the system is
| + | <div class="container"> |
− | interrelated by metabolic or regulatory pathways. Integrated with the systems, the Pichia
| + | <h1 id="Top">Proof Of Concept</h1> |
− | pastoris GS115 can express target products which are separated and applied with our hardware. We
| + | <p class="lead pl-1"></p> |
− | will prove the validity and feasibility from the following three aspects including construction,
| + | <hr class="my-4" /> |
− | expression and application.</p>
| + | </div> |
− | <h1>Construction</h1>
| + | </header> |
− | <div class="image"><img alt=" "
| + | <main> |
− | src="https://static.igem.org/mediawiki/2021/d/d7/T--HUST-China--img--proof--proof1.png"
| + | <div class="container"> |
− | style="width: 100%"/>
| + | <div class="row"> |
| + | <div class="sidebar col-lg-3"> |
| + | <div class="nav" id="contents"> |
| + | <h5>Contents</h5> |
| + | <ul></ul> |
| </div> | | </div> |
− | <p>Every system is composed of dye and perm device as well as decolorization and straightening | + | </div> |
− | device, constituted of several parts. The construction process shall be divided into two parts:
| + | <div class="content col-lg-9"> |
− | first connecting the parts to build devices, next assembling devices into systems. During this
| + | <article> |
− | process, we chose seamless cloning of homologous recombination at the beginning and turned to
| + | <h1>Introduction</h1> |
− | enzyme digestion and ligation, finally getting the expected result through the latter
| + | <p>Our project (click <a |
− | method.</p>
| + | href="https://2021.igem.org/Team:HUST-China/Design">design</a> for |
− | <div class="modal-btn">
| + | detailed |
− | <button class="btn btn-warning pull-right" data-target="#Proof1" data-toggle="modal"
| + | information) integrated at most five reactions within a system. Each reaction in the system |
| + | is |
| + | interrelated by metabolic or regulatory pathways. Integrated with the systems, the Pichia |
| + | pastoris GS115 can express target products which are separated and applied with our |
| + | hardware. We |
| + | will prove the validity and feasibility from the following three aspects including |
| + | construction, |
| + | expression and application.</p> |
| + | <h1>Construction</h1> |
| + | <div class="image"><img alt=" " |
| + | src="https://static.igem.org/mediawiki/2021/d/d7/T--HUST-China--img--proof--proof1.png" |
| + | style="width: 100%" /> |
| + | </div> |
| + | <p>Every system is composed of dye and perm device as well as decolorization and straightening |
| + | device, constituted of several parts. The construction process shall be divided into two |
| + | parts: |
| + | first connecting the parts to build devices, next assembling devices into systems. During |
| + | this |
| + | process, we chose seamless cloning of homologous recombination at the beginning and turned |
| + | to |
| + | enzyme digestion and ligation, finally getting the expected result through the latter |
| + | method.</p> |
| + | <div class="modal-btn"> |
| + | <button class="btn btn-warning pull-right" data-target="#Proof1" data-toggle="modal" |
| type="button">Read More | | type="button">Read More |
− | </button>
| + | </button> |
− | <div aria-hidden="true" aria-labelledby="Proof1Label" class="modal fade" data-backdrop="static"
| + | <div aria-hidden="true" aria-labelledby="Proof1Label" class="modal fade" |
− | data-keyboard="false" id="Proof1" tabindex="-1">
| + | data-backdrop="static" data-keyboard="false" id="Proof1" tabindex="-1"> |
− | <div class="modal-dialog modal-dialog-scrollable modal-dialog-centered modal-xl">
| + | <div class="modal-dialog modal-dialog-scrollable modal-dialog-centered modal-xl"> |
− | <div class="modal-content">
| + | <div class="modal-content"> |
− | <div class="modal-header"><h5 class="modal-title" id="Proof1Label">Proof1</h5></div>
| + | <div class="modal-header"> |
− | <div class="modal-body"><p>In the initial stage of construction, we adapted seamless cloning of homologous recombination.
| + | <h5 class="modal-title" id="Proof1Label">Proof1</h5> |
− | Unique homologous sequences at front and rear ends of joining parts were designed based on
| + | </div> |
− | Gibson assembly seamless cloning principle. After PCR, the parts underwent recombination
| + | <div class="modal-body"> |
− | reaction in the recombination enzyme system from some companies. Unfortunately, the products of
| + | <p>In the initial stage of construction, we adapted seamless cloning of |
− | ligation fell short of our expectation and we hardly found any explanation to this. Later, the
| + | homologous recombination. |
− | restricted enzyme and T4 ligation enzyme were used to construct the devices, which is a less
| + | Unique homologous sequences at front and rear ends of joining parts were |
− | efficient way of construction. Most devices and a few systems were successfully constructed and
| + | designed based on |
− | we will continue to complete our work in the future. More details please click <a
| + | Gibson assembly seamless cloning principle. After PCR, the parts |
− | href="https://2021.igem.org/Team:HUST-China/Proof_Of_Concept/Engineering">here</a></p></div>
| + | underwent recombination |
− | <div class="modal-footer">
| + | reaction in the recombination enzyme system from some companies. |
− | <button class="btn btn-danger" data-dismiss="modal" type="button">Close</button>
| + | Unfortunately, the products of |
| + | ligation fell short of our expectation and we hardly found any |
| + | explanation to this. Later, the |
| + | restricted enzyme and T4 ligation enzyme were used to construct the |
| + | devices, which is a less |
| + | efficient way of construction. Most devices and a few systems were |
| + | successfully constructed and |
| + | we will continue to complete our work in the future. More details please |
| + | click <a |
| + | href="https://2021.igem.org/Team:HUST-China/Engineering">here</a> |
| + | </p> |
| + | </div> |
| + | <div class="modal-footer"> |
| + | <button class="btn btn-danger" data-dismiss="modal" |
| + | type="button">Close</button> |
| + | </div> |
| </div> | | </div> |
| </div> | | </div> |
| </div> | | </div> |
| </div> | | </div> |
− | </div>
| + | <h1>Expression</h1> |
− | <h1>Expression</h1>
| + | <p>The constructed devices and systems were transformed into pichia pastoris GS115 and |
− | <p>The constructed devices and systems were transformed into pichia pastoris GS115 and expressed. We
| + | expressed. We |
− | added α-factor as the signal peptide in front of the functional genes for extra-cellular
| + | added α-factor as the signal peptide in front of the functional genes for extra-cellular |
− | secretion but ended up unsatisfactory. The signal peptide was removed for intracellular
| + | secretion but ended up unsatisfactory. The signal peptide was removed for intracellular |
− | expression and we got ideal result, with most protein successfully expressed. The activity of
| + | expression and we got ideal result, with most protein successfully expressed. The activity |
− | two kinds of enzymes were measured.</p>
| + | of |
− | <div class="modal-btn">
| + | two kinds of enzymes were measured.</p> |
− | <button class="btn btn-warning pull-right" data-target="#Proof2" data-toggle="modal"
| + | <div class="modal-btn"> |
| + | <button class="btn btn-warning pull-right" data-target="#Proof2" data-toggle="modal" |
| type="button">Read More | | type="button">Read More |
− | </button>
| + | </button> |
− | <div aria-hidden="true" aria-labelledby="Proof2Label" class="modal fade" data-backdrop="static"
| + | <div aria-hidden="true" aria-labelledby="Proof2Label" class="modal fade" |
− | data-keyboard="false" id="Proof2" tabindex="-1">
| + | data-backdrop="static" data-keyboard="false" id="Proof2" tabindex="-1"> |
− | <div class="modal-dialog modal-dialog-scrollable modal-dialog-centered modal-xl">
| + | <div class="modal-dialog modal-dialog-scrollable modal-dialog-centered modal-xl"> |
− | <div class="modal-content">
| + | <div class="modal-content"> |
− | <div class="modal-header"><h5 class="modal-title" id="Proof2Label">Proof2</h5></div>
| + | <div class="modal-header"> |
− | <div class="modal-body"><h1>Chassis Yeast</h1>
| + | <h5 class="modal-title" id="Proof2Label">Proof2</h5> |
− | <p>Pichia pastoris, a kind of methylotrophic yeast separated by Ogata in 1969, can grow in culture
| + | </div> |
− | media with methanol as the only carbon source.[1].P.pastoris, being the second widely used
| + | <div class="modal-body"> |
− | exogenous protein expression system, is widely applied in expression of exogenous protein.
| + | <h2>Chassis Yeast</h2> |
− | Pichia pastoris expression system has good post-translational modification function and
| + | <p>Pichia pastoris, a kind of methylotrophic yeast separated by Ogata in |
− | secretion efficiency. Compared with Saccharomyces cerevisiae expression system, its
| + | 1969, can grow in culture |
− | glycosylation degree is low and can be cultured in high density. After sequencing the whole
| + | media with methanol as the only carbon source.[1].P.pastoris, being the |
− | genome of Pichia pastoris in 2009[2], its genetic background became clear, which further
| + | second widely used |
− | promoted the research of Pichia pastoris expression system. At present, more than 5000 exogenous
| + | exogenous protein expression system, is widely applied in expression of |
− | proteins have been successfully expressed in P. pastoris, and P. pastoris has become one of the
| + | exogenous protein. |
− | most commonly used expression systems.[3].</p>
| + | Pichia pastoris expression system has good post-translational |
− | <h1>Xylose Responding Regulation System</h1>
| + | modification function and |
− | <p>In order to express the genes at the right time, we introduced a Xylose Responding Regulation
| + | secretion efficiency. Compared with Saccharomyces cerevisiae expression |
− | System. The system includes two promoters: the constitutive promoter Panb1, applied in the dye
| + | system, its |
− | and perm devices and could be inhibited by repressor ROX1[4]; and the xylose induced promoter
| + | glycosylation degree is low and can be cultured in high density. After |
− | Pynr071c[5], applied in the decolorization and straightening devices. And the two promoters have
| + | sequencing the whole |
− | been proved that would express in Saccharomyces cerevisiae, so we try them in P.pastoris.</p>
| + | genome of Pichia pastoris in 2009[2], its genetic background became |
− | <h1>Expression of parts</h1>
| + | clear, which further |
− | <p>As for our functional genes, they had been expressed in some chassis. FMO <a
| + | promoted the research of Pichia pastoris expression system. At present, |
− | href="http://parts.igem.org/Part:BBa_K3711003">BBa_K3711003</a> in E.coli, Laccase <a
| + | more than 5000 exogenous |
− | href="http://parts.igem.org/Part:BBa_K3711004">BBa_K3711004</a> in P.pastoris[6], crtE <a
| + | proteins have been successfully expressed in P. pastoris, and P. |
− | href="http://parts.igem.org/Part:BBa_K3711005">BBa_K3711005</a>, crtB <a
| + | pastoris has become one of the |
− | href="http://parts.igem.org/Part:BBa_K3711006">BBa_K3711006</a>, crtI <a
| + | most commonly used expression systems.[3].</p> |
− | href="http://parts.igem.org/Part:BBa_K3711006">BBa_K3711006</a> in S. cerevisiae[7], 4CL <a
| + | <h2>Xylose Responding Regulation System</h2> |
− | href="http://parts.igem.org/Part:BBa_K3711009">BBa_K3711009</a>, ACC <a
| + | <p>In order to express the genes at the right time, we introduced a Xylose |
− | href="http://parts.igem.org/Part:BBa_K3711010">BBa_K3711010</a>, CUS <a
| + | Responding Regulation |
− | href="http://parts.igem.org/Part:BBa_K3711011">BBa_K3711011</a> in E.coli, curA <a
| + | System. The system includes two promoters: the constitutive promoter |
− | href="http://parts.igem.org/Part:BBa_K3711012">BBa_K3711012</a> in P.pastoris[8]. So we try
| + | Panb1, applied in the dye |
− | to express them in eukaryotic system P.pastoris GS115.</p></div>
| + | and perm devices and could be inhibited by repressor ROX1[4]; and the |
− | <div class="modal-footer">
| + | xylose induced promoter |
− | <button class="btn btn-danger" data-dismiss="modal" type="button">Close</button>
| + | Pynr071c[5], applied in the decolorization and straightening devices. |
| + | And the two promoters have |
| + | been proved that would express in Saccharomyces cerevisiae, so we try |
| + | them in P.pastoris.</p> |
| + | <h2>Expression of parts</h2> |
| + | <p>As for our functional genes, they had been expressed in some chassis. FMO |
| + | <a href="http://parts.igem.org/Part:BBa_K3711003">BBa_K3711003</a> in |
| + | E.coli, Laccase <a |
| + | href="http://parts.igem.org/Part:BBa_K3711004">BBa_K3711004</a> in |
| + | P.pastoris[6], crtE <a |
| + | href="http://parts.igem.org/Part:BBa_K3711005">BBa_K3711005</a>, |
| + | crtB <a href="http://parts.igem.org/Part:BBa_K3711006">BBa_K3711006</a>, |
| + | crtI <a href="http://parts.igem.org/Part:BBa_K3711006">BBa_K3711006</a> |
| + | in S. cerevisiae[7], 4CL <a |
| + | href="http://parts.igem.org/Part:BBa_K3711009">BBa_K3711009</a>, ACC |
| + | <a href="http://parts.igem.org/Part:BBa_K3711010">BBa_K3711010</a>, CUS |
| + | <a href="http://parts.igem.org/Part:BBa_K3711011">BBa_K3711011</a> in |
| + | E.coli, curA <a |
| + | href="http://parts.igem.org/Part:BBa_K3711012">BBa_K3711012</a> in |
| + | P.pastoris[8]. So we try |
| + | to express them in eukaryotic system P.pastoris GS115.</p> |
| + | </div> |
| + | <div class="modal-footer"> |
| + | <button class="btn btn-danger" data-dismiss="modal" |
| + | type="button">Close</button> |
| + | </div> |
| </div> | | </div> |
| </div> | | </div> |
| </div> | | </div> |
| </div> | | </div> |
− | </div>
| + | <h1>Application</h1> |
− | <h1>Application</h1>
| + | <p>When it comes to the application of our products, we should ensure the theoretical basis of |
− | <p>When it comes to the application of our products, we should ensure the theoretical basis of our
| + | our |
− | pathways. Furthermore, to put it into the real world for use, we need the support of hardware.
| + | pathways. Furthermore, to put it into the real world for use, we need the support of |
− | In this section, we introduce these two aspects.</p>
| + | hardware. |
− | <div class="modal-btn">
| + | In this section, we introduce these two aspects.</p> |
− | <button class="btn btn-warning pull-right" data-target="#Proof3" data-toggle="modal"
| + | <div class="modal-btn"> |
| + | <button class="btn btn-warning pull-right" data-target="#Proof3" data-toggle="modal" |
| type="button">Read More | | type="button">Read More |
− | </button>
| + | </button> |
− | <div aria-hidden="true" aria-labelledby="Proof3Label" class="modal fade" data-backdrop="static"
| + | <div aria-hidden="true" aria-labelledby="Proof3Label" class="modal fade" |
− | data-keyboard="false" id="Proof3" tabindex="-1">
| + | data-backdrop="static" data-keyboard="false" id="Proof3" tabindex="-1"> |
− | <div class="modal-dialog modal-dialog-scrollable modal-dialog-centered modal-xl">
| + | <div class="modal-dialog modal-dialog-scrollable modal-dialog-centered modal-xl"> |
− | <div class="modal-content">
| + | <div class="modal-content"> |
− | <div class="modal-header"><h5 class="modal-title" id="Proof3Label">Proof3</h5></div>
| + | <div class="modal-header"> |
− | <div class="modal-body"><h1>The pigments can be used to dye</h1>
| + | <h5 class="modal-title" id="Proof3Label">Proof3</h5> |
− | <div class="image"><img alt=" "
| + | </div> |
− | src="https://static.igem.org/mediawiki/2021/d/d7/T--HUST-China--img--proof--proof2.png"
| + | <div class="modal-body"> |
− | style="width: 100%"/>
| + | <h2>The pigments can be used to dye</h2> |
− | </div>
| + | <div class="image"><img alt=" " |
− | <p>Before our experiments, indigo and curcumin had been researched in dye[9][10].Then we had
| + | src="https://static.igem.org/mediawiki/2021/b/b5/T--HUST-China--Proof_2.jpeg" |
− | researched a lot about dye with the three pigments. We dyed the hair successfully and compared
| + | style="width: 60%" /> |
− | it with the hair dyed with the traditional chemical way. Please refer to <a
| + | </div> |
− | href="https://2021.igem.org/Team:HUST-China/Proof_Of_Concept/Results">result</a> for
| + | <p>Before our experiments, indigo and curcumin had been researched in |
− | more information.</p>
| + | dye[9][10].Then we had |
− | <h1>The Color of Hairs Can Be Faded</h1>
| + | researched a lot about dye with the three pigments. We dyed the hair |
− | <p>In our design, we have gained information from papers that lipoxygenase can fade the color
| + | successfully and compared |
− | produced by lycopene[11], curcumin reductase can fade curcumin[12], and indigo can be faded by
| + | it with the hair dyed with the traditional chemical way. Please refer to |
− | laccase[13]. We also finished some experiments to prove these enzymes can work as we thought. <a
| + | <a |
− | href="https://2021.igem.org/Team:HUST-China/Proof_Of_Concept/Results">(Results)</a></p>
| + | href="https://2021.igem.org/Team:HUST-China/Results">result</a> |
− | <h1>The Short Peptides Can Be Used to Perm</h1>
| + | for |
− | <div class="image"><img alt=" "
| + | more information.</p> |
− | src="https://static.igem.org/mediawiki/2021/a/ab/T--HUST-China--img--proof--proof3.png"
| + | <h2>The Color of Hairs Can Be Faded</h2> |
− | style="width: 100%"/>
| + | <p>In our design, we have gained information from papers that lipoxygenase |
− | </div>
| + | can fade the color |
− | <p>We have known that traditional perming ways use strong allergen to open the cortex of hairs. To
| + | produced by lycopene[11], curcumin reductase can fade curcumin[12], and |
− | change and improve this, we designed a new short peptide PepACS, consisting of PepA, PepC and a
| + | indigo can be faded by |
− | human lung protein surfactant B called SPB, as calculated and optimized by modeling. The peptide
| + | laccase[13]. We also finished some experiments to prove these enzymes |
− | is small enough to enter the cortex through the cuticle and form re-conformed disulfide bonds
| + | can work as we thought. <a |
− | with α-keratins, thus changing the shape of hair[14]. We also designed some experiments to prove
| + | href="https://2021.igem.org/Team:HUST-China/Results">(Results)</a> |
− | it and more information please refer to <a
| + | </p> |
− | href="https://2021.igem.org/Team:HUST-China/Proof_Of_Concept/Results">result</a>.</p>
| + | <h2>The Short Peptides Can Be Used to Perm</h2> |
− | <h1>Use Our Products in Real Life</h1>
| + | <div class="image"><img alt=" " |
− | <p>For the use of our products in reality, we have designed two main apparatuses. The fermentation
| + | src="https://static.igem.org/mediawiki/2021/6/6f/T--HUST-China--Proof_3.png" |
− | bottle is used to produce pigments and short peptides. And the dyeing comb is used to change the
| + | style="width: 40%" /> |
− | color and shape of your hairs. With the help of them, you can finish the process of perming and
| + | </div> |
− | dyeing. Please refer to <a
| + | <p>We have known that traditional perming ways use strong allergen to open |
− | href="https://2021.igem.org/Team:HUST-China/Proof_Of_Concept/Hardware">Hardware</a> for
| + | the cortex of hairs. To |
− | more information.</p></div>
| + | change and improve this, we designed a new short peptide PepACS, |
− | <div class="modal-footer">
| + | consisting of PepA, PepC and a |
− | <button class="btn btn-danger" data-dismiss="modal" type="button">Close</button>
| + | human lung protein surfactant B called SPB, as calculated and optimized |
| + | by modeling. The peptide |
| + | is small enough to enter the cortex through the cuticle and form |
| + | re-conformed disulfide bonds |
| + | with α-keratins, thus changing the shape of hair[14]. We also designed |
| + | some experiments to prove |
| + | it and more information please refer to <a |
| + | href="https://2021.igem.org/Team:HUST-China/Results">result</a>. |
| + | </p> |
| + | <h2>Use Our Products in Real Life</h2> |
| + | <p>For the use of our products in reality, we have designed two main |
| + | apparatuses. The fermentation |
| + | bottle is used to produce pigments and short peptides. And the dyeing |
| + | comb is used to change the |
| + | color and shape of your hairs. With the help of them, you can finish the |
| + | process of perming and |
| + | dyeing. Please refer to <a |
| + | href="https://2021.igem.org/Team:HUST-China/Hardware">Hardware</a> |
| + | for |
| + | more information.</p> |
| + | </div> |
| + | <div class="modal-footer"> |
| + | <button class="btn btn-danger" data-dismiss="modal" |
| + | type="button">Close</button> |
| + | </div> |
| </div> | | </div> |
| </div> | | </div> |
| </div> | | </div> |
| </div> | | </div> |
− | </div>
| |
| | | |
− | <div class="row g-0">
| + | <div class="row g-0"> |
− | <div class="col-4 girl"><img
| + | <div class="col-4 girl"><img |
− | src="https://static.igem.org/mediawiki/2021/2/28/T--HUST-China--img--girl.png"/></div>
| + | src="https://static.igem.org/mediawiki/2021/2/28/T--HUST-China--img--girl.png" /></div> |
− | <div class="col-5">
| + | <div class="col-5"> |
− | <div class="dialogbox"><p>Hmm…I can hardly pick out any flaw in this part, what about you?
| + | <div class="dialogbox"> |
− | Here is another thing I really care about—the <a
| + | <p>Hmm…I can hardly pick out any flaw in this part, what about you? |
− | href="https://2021.igem.org/Team:HUST-China/Proof_Of_Concept/Safety">safety </a>of
| + | Here is another thing I really care about—the <a |
− | the products! Don’t you worry about whether the products of this team can cause cancer
| + | href="https://2021.igem.org/Team:HUST-China/Safety">safety |
− | or do damage to hair like other hairdressing products in the market?</p></div>
| + | </a>of |
| + | the products! Don’t you worry about whether the products of this team can cause |
| + | cancer |
| + | or do damage to hair like other hairdressing products in the market?</p> |
| + | </div> |
| + | </div> |
| </div> | | </div> |
− | </div> | + | </article> |
− | </article> | + | </div> |
| </div> | | </div> |
| </div> | | </div> |
− | </div> | + | </main> |
− | </main> | + | <footer> |
− | <footer> | + | <div class="container"> |
− | <div class="container">
| + | <div class="row"> |
− | <div class="row">
| + | <div class="col-7"> |
− | <div class="col-7">
| + | <div class="row justify-content-center"> |
− | <div class="row justify-content-center">
| + | <div class="footer-logo"></div> |
− | <div class="footer-logo"></div>
| + | </div> |
| + | <p><span>CONTACT US: </span><a href="mailto:iGEMHUSTChina@163.com">iGEMHUSTChina@163.com</a></p> |
| + | <p>Huazhong University of Sci. & Tech., Wuhan, China</p> |
| + | <p>1037# Luoyu Rd, Wuhan, P.R.China 430074</p> |
| + | <p>Copyright © <span>HUST-China </span>iGEM 2021</p> |
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− | <p><span>CONTACT US: </span><a href="mailto:iGEMHUSTChina@163.com">iGEMHUSTChina@163.com</a></p> | + | <div class="col-5"> |
− | <p>Huazhong University of Sci. & Tech., Wuhan, China</p>
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− | <p>1037# Luoyu Rd, Wuhan, P.R.China 430074</p>
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− | <p>Copyright © <span>HUST-China </span>iGEM 2021</p></div>
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