Difference between revisions of "Team:Shanghai Metro HS/Results"
| Line 96: | Line 96: | ||
<div class="sub-content"> | <div class="sub-content"> | ||
<div class="img-wrap no-margin"> | <div class="img-wrap no-margin"> | ||
| − | <img src="https://static.igem.org/mediawiki/2021/d/d6/T--Shanghai_Metro_HS--results01.jpg" alt="" /> | + | <img tyle="width: 816px;" src="https://static.igem.org/mediawiki/2021/d/d6/T--Shanghai_Metro_HS--results01.jpg" |
| + | alt="" /> | ||
<span>Figure 1. The result of enzyme cutting and PCR</span> | <span>Figure 1. The result of enzyme cutting and PCR</span> | ||
</div> | </div> | ||
| − | <div class="article-content">This picture is the nucleic acid electrophoresis result of enzyme cutting and | + | <div class="article-content" style="font-size: 18px;">This picture is the nucleic acid electrophoresis result of |
| + | enzyme cutting and | ||
PCR(By performing PCR, we can obtain the desirable PKC-OP’s gene segment.). Column “marker” is a column that | PCR(By performing PCR, we can obtain the desirable PKC-OP’s gene segment.). Column “marker” is a column that | ||
is used to show the position of different lengths of genes. In this step, our aim is to verify whether these | is used to show the position of different lengths of genes. In this step, our aim is to verify whether these | ||
| Line 109: | Line 111: | ||
</div> | </div> | ||
<div class="img-wrap no-margin"> | <div class="img-wrap no-margin"> | ||
| − | <img src="https://static.igem.org/mediawiki/2021/6/61/T--Shanghai_Metro_HS--results02.jpg" alt="" /> | + | <img tyle="width: 816px;" src="https://static.igem.org/mediawiki/2021/6/61/T--Shanghai_Metro_HS--results02.jpg" |
| + | alt="" /> | ||
<span>Figure 2 E. coil having the desired pET-25b and PKC-OP </span> | <span>Figure 2 E. coil having the desired pET-25b and PKC-OP </span> | ||
</div> | </div> | ||
<div class="article-content">The pET25b-PKC-OP was constructed.</div> | <div class="article-content">The pET25b-PKC-OP was constructed.</div> | ||
| − | <div class="article-content">The plate shows monoclonals of pET25b-PKC-OP constructs </div> | + | <div class="article-content" style="font-size: 18px;">The plate shows monoclonals of pET25b-PKC-OP constructs |
| + | </div> | ||
<div class="img-wrap no-margin"> | <div class="img-wrap no-margin"> | ||
| − | <img src="https://static.igem.org/mediawiki/2021/b/b3/T--Shanghai_Metro_HS--results03.jpg" alt="" /> | + | <img tyle="width: 816px;" src="https://static.igem.org/mediawiki/2021/b/b3/T--Shanghai_Metro_HS--results03.jpg" |
| + | alt="" /> | ||
<span>Figure 3. The result of Apa1 enzyme digestion validation</span> | <span>Figure 3. The result of Apa1 enzyme digestion validation</span> | ||
</div> | </div> | ||
| − | <div class="article-content"> | + | <div class="article-content" style="font-size: 18px;"> |
Column “marker” is a column that is used to show the position of different lengths of genes. Number 1 to 10 | Column “marker” is a column that is used to show the position of different lengths of genes. Number 1 to 10 | ||
is the result for recombinant plasmid pET25b-PKC-OP after Apa1 enzyme digestion. We get two bands of 4712bp | is the result for recombinant plasmid pET25b-PKC-OP after Apa1 enzyme digestion. We get two bands of 4712bp | ||
and 1894bp. It further indicates that the obtained monoclonals were positive monoclonals containing the | and 1894bp. It further indicates that the obtained monoclonals were positive monoclonals containing the | ||
recombinant plasmid.</div> | recombinant plasmid.</div> | ||
| − | <div class="article-content">Future plan: Testing the effectiveness of producing cellulase of positive | + | <div class="article-content" style="font-size: 18px;">Future plan: Testing the effectiveness of producing |
| + | cellulase of positive | ||
recombinant bacteria. This is important because we need to know how much bacteria is needed to be used to | recombinant bacteria. This is important because we need to know how much bacteria is needed to be used to | ||
effectively help animals digest fodder. In this experiment, we can control the time that positive | effectively help animals digest fodder. In this experiment, we can control the time that positive | ||
| Line 130: | Line 136: | ||
</div> | </div> | ||
<div class="img-wrap no-margin"> | <div class="img-wrap no-margin"> | ||
| − | <img src="https://static.igem.org/mediawiki/2021/5/55/T--Shanghai_Metro_HS--chgt3.jpg" alt="" /> | + | <img style="width: 816px;" src="https://static.igem.org/mediawiki/2021/5/55/T--Shanghai_Metro_HS--chgt3.jpg" |
| + | alt="" /> | ||
</div> | </div> | ||
<div class="img-wrap no-margin"> | <div class="img-wrap no-margin"> | ||
| − | <img src="https://static.igem.org/mediawiki/2021/7/78/T--Shanghai_Metro_HS--chgt4.jpg" alt="" /> | + | <img style="width: 816px;" src="https://static.igem.org/mediawiki/2021/7/78/T--Shanghai_Metro_HS--chgt4.jpg" |
| + | alt="" /> | ||
</div> | </div> | ||
| − | <div class="article-content"> | + | <div class="article-content" style="font-size: 18px;"> |
We designed several different induction conditions for protein expression. As the vector we chose to | We designed several different induction conditions for protein expression. As the vector we chose to | ||
construct the plasmid contains a pelB signal peptide, the engineered strain would secret the protein into | construct the plasmid contains a pelB signal peptide, the engineered strain would secret the protein into | ||
| Line 145: | Line 153: | ||
</div> | </div> | ||
<div class="article-title">Future Plan </div> | <div class="article-title">Future Plan </div> | ||
| − | <div class="article-content">Testing the effectiveness of producing cellulase of positive recombinant bacteria. | + | <div class="article-content" style="font-size: 18px;">Testing the effectiveness of producing cellulase of |
| + | positive recombinant bacteria. | ||
This is important because we need to know how much bacteria is needed to be used to effectively help animals | This is important because we need to know how much bacteria is needed to be used to effectively help animals | ||
digest fodder. In this experiment, we can control the time that positive recombinant bacteria grow and use | digest fodder. In this experiment, we can control the time that positive recombinant bacteria grow and use | ||
Latest revision as of 13:02, 20 October 2021
Results
Figure 1. The result of enzyme cutting and PCR
This picture is the nucleic acid electrophoresis result of
enzyme cutting and
PCR(By performing PCR, we can obtain the desirable PKC-OP’s gene segment.). Column “marker” is a column that
is used to show the position of different lengths of genes. In this step, our aim is to verify whether these
results are the desired ones.
In figure 1, the number labeled above figure 1 represents different results. To begin with, number 1 and 2
is the result for pET-25b after enzyme cutting. Additionally, number 3 is the pET-25b without enzyme
cutting. At last, number 4 and 5 is the PKC-OP after PCR. As our experiment moves on, the data of our PKC-OP
and pET-25b are 233.2ng/μL and 17.8ng/μL. Lastly, we use the homologous combination to combine them.
Figure 2 E. coil having the desired pET-25b and PKC-OP
The pET25b-PKC-OP was constructed.
The plate shows monoclonals of pET25b-PKC-OP constructs
Figure 3. The result of Apa1 enzyme digestion validation
Column “marker” is a column that is used to show the position of different lengths of genes. Number 1 to 10
is the result for recombinant plasmid pET25b-PKC-OP after Apa1 enzyme digestion. We get two bands of 4712bp
and 1894bp. It further indicates that the obtained monoclonals were positive monoclonals containing the
recombinant plasmid.
Future plan: Testing the effectiveness of producing
cellulase of positive
recombinant bacteria. This is important because we need to know how much bacteria is needed to be used to
effectively help animals digest fodder. In this experiment, we can control the time that positive
recombinant bacteria grow and use the OD test to set the numbers of bacteria. Then put different numbers of
bacteria and cellulose filter paper together. OD test can be used again to see the glucose concentration.
We designed several different induction conditions for protein expression. As the vector we chose to
construct the plasmid contains a pelB signal peptide, the engineered strain would secret the protein into
the medium. Therefore, we collected the culture supernatant after induction and ran SDS-PAGE for
verification.
According to the calculation by Snapgene, the expected protein molecular shall weigh 45.6 KDa. As seen from
the SDS-PAGE map (Fig. 4), there is a wide band just above 40 KDa and it indicates that we have obtained the
PKC-OP protein.
Future Plan
Testing the effectiveness of producing cellulase of
positive recombinant bacteria.
This is important because we need to know how much bacteria is needed to be used to effectively help animals
digest fodder. In this experiment, we can control the time that positive recombinant bacteria grow and use
the OD test to set the numbers of bacteria. Then put different numbers of bacteria and cellulose filter
paper together. OD test can be used again to see the glucose concentration.