Team:HUST-China/Parts

Parts | iGEM HUST-China

Parts


Introduction

Biology is different from other engineering fields due to a lack of modular nature of the structure and function, which means the expression of gene will change when transferred from one species to another. What synthetic biology is dedicated to is the modularization and standardization of functional gene, to make the expression of it predictable in different chassis organism. Our project has registered 19 basic parts and 56 composite parts, all qualified for standardized requirement of RFC[10]. We have also tested these parts. We hope that everyone can utilize our parts based on our research through registry and that the information for parts can further be completed so that more people can make use of them.

Basic parts

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Part Number Part Name Type length
BBa_K3711019 Panb1 Regulatory 399
BBa_K3711001 Pynr071c Regulatory 1188
BBa_K3711015 AOX1 Regulatory 937
BBa_K3711017 AOX1 terminator Terminator 247
BBa_K3711002 ROX1 Coding 1251
BBa_K3711003 FMO Coding 1395
BBa_K3711004 Laccase Coding 1602
BBa_K3711005 crtE Coding 927
BBa_K3711006 crtB Coding 948
BBa_K3711007 crtI Coding 1497
BBa_K3711008 LOX2 Coding 2709
BBa_K3711009 4CL Coding 1704
BBa_K3711010 ACC Coding 1800
BBa_K3711011 CUS Coding 1227
BBa_K3711012 curA Coding 1056
BBa_K3711013 PepACS Coding 168
BBa_K3711014 DsbC Coding 903
BBa_K3711016 α-factor Coding 267
BBa_K3711018 FMO dimer Coding 2769

Improvement

Flaving Containing-Monooxygenase (FMO) is an enzyme that can be used to produce indigo. This basic part has been registered by iGEM13_Berkeley as BBa_K1131000. We improved its function and registered as a new part BBa_K3711018.

Firstly, we optimized the codon of this gene according to the codon preference of P.pastoris to improve its expression level.

Secondly, research shows that FAD is the prosthetic group of FMO, NADPH is the cofactor of FMO. FMO-FAD-NADPH complex usually forms a asymmetric dimer as a unit to work. The enzyme-substrate complex formed when enzyme acts on substrate, has two dimers per unit[1].

So, we wanted to construct the two FMO fusion proteins named FMO dimer and expressed it, hoping that they could form a dimer more easily and improve their activity.

Then, we predicted the 3D structure of FMO dimer. The result shows that the structure could be formed and the dimer could combinate with substrate.

Therefore, we started to build FMO dimer and achieved success in expression. More details please click here.

Composite parts

Part Number Part Name Type length
BBa_K3711039 AOX1-α factor-ROX1-AOX1 Terminator Composite 2722
BBa_K3711040 AOX1-α factor-FMO-AOX1 Terminator Composite 2866
BBa_K3711041 AOX1-α factor-Laccase-AOX1 Terminator Composite 3073
BBa_K3711042 AOX1-α factor-crtE-AOX1 Terminator Composite 2398
BBa_K3711043 AOX1-α factor-crtB-AOX1 Terminator Composite 2419
BBa_K3711044 AOX1-α factor-crtI-AOX1 Terminator Composite 2968
BBa_K3711045 AOX1-α factor-LOX2-AOX1 Terminator Composite 4180
BBa_K3711046 AOX1-α factor-4CL-AOX1 Terminator Composite 3175
BBa_K3711047 AOX1-α factor-ACC-AOX1 Terminator Composite 3271
BBa_K3711048 AOX1-α factor-CUS-AOX1 Terminator Composite 2698
BBa_K3711049 AOX1-α factor-curA-AOX1 Terminator Composite 2527
BBa_K3711050 AOX1-α factor-PepACS-AOX1 Terminator Composite 1639
BBa_K3711051 AOX1-α factor-DsbC-AOX1 Terminator Composite 2374
BBa_K3711052 Pynr071c-α factor-FMO dimer-AOX1 Terminator Composite 4491
BBa_K3711053 Pynr071c-α factor-ROX1-AOX1 Terminator Composite 2973
BBa_K3711054 Panb1-α factor-FMO-AOX1 Terminator Composite 2328
BBa_K3711055 Pynr071c-α factor-Laccase-AOX1 Terminator Composite 3324
BBa_K3711056 Panb1-α factor-crtE-AOX1 Terminator Composite 1860
BBa_K3711057 Panb1-α factor-crtB-AOX1 Terminator Composite 1881
BBa_K3711058 Panb1-α factor-crtI-AOX1 Terminator Composite 2430
BBa_K3711060 Pynr071c-α factor-LOX2-AOX1 Terminator Composite 4431
BBa_K3711061 Panb1-α factor-4CL-AOX1 Terminator Composite 2637
BBa_K3711062 Panb1-α factor-ACC-AOX1 Terminator Composite 2733
BBa_K3711063 Panb1-α factor-CUS-AOX1 Terminator Composite 2160
BBa_K3711064 Pynr071c-α factor-curA-AOX1 Terminator Composite 2778
BBa_K3711065 Panb1-α factor-PepACS-AOX1 Terminator Composite 1101
BBa_K3711066 Pynr071c-α factor-DsbC-AOX1 Terminator Composite 2625
BBa_K3711067 Pynr071c-ROX1-AOX1 Terminator Composite 2700
BBa_K3711068 Panb1-FMO-AOX1 Terminator Composite 2055
BBa_K3711069 Pynr071c-Laccase-AOX1 Terminator Composite 3051
BBa_K3711070 Panb1-crtE-AOX1 Terminator Composite 1587
BBa_K3711071 Panb1-crtB-AOX1 Terminator Composite 1608
BBa_K3711072 Panb1-crtI-AOX1 Terminator Composite 2157
BBa_K3711073 Pynr071c-LOX2-AOX1 Terminator Composite 4158
BBa_K3711074 Panb1-4CL-AOX1 Terminator Composite 2364
BBa_K3711075 Panb1-ACC-AOX1 Terminator Composite 2460
BBa_K3711076 Panb1-CUS-AOX1 Terminator Composite 1887
BBa_K3711077 Pynr071c-curA-AOX1 Terminator Composite 2505
BBa_K3711078 Panb1-PepACS-AOX1 Terminator Composite 828
BBa_K3711079 Pynr071c-DsbC-AOX1 Terminator Composite 2352
BBa_K3711080 AOX1-α factor-Lycopene-AOX1 Terminator Composite 7801
BBa_K3711081 AOX1-α factor-Curcumin-AOX1 Terminator Composite 9160
BBa_K3711082 Panb1-α factor-Curcumin-AOX1 Terminator Composite 7546
BBa_K3711083 Panb1-α factor-Lycopene-AOX1 Terminator Composite 6187
BBa_K3711084 Panb1-Curcumin-AOX1 Terminator Composite 6727
BBa_K3711085 Panb1-Lycopene-AOX1 Terminator Composite 5368
BBa_K3711086 The synthesis and decolorization system of indigo Composite 8368
BBa_K3711087 The synthesis and decolorization system of indigo Composite 7822
BBa_K3711088 The synthesis and decolorization system of indigo Composite 14948
BBa_K3711089 The synthesis and decolorization system of lycopene without signal peptide Composite 12242
BBa_K3711090 The synthesis and decolorization system of lycopene without signal peptide Composite 13040
BBa_K3711091 The synthesis and decolorization system of curcumin without signal peptide Composite 11948
BBa_K3711092 The perm and straightening system Composite 6442
BBa_K3711093 The perm and straightening system without signal peptide Composite 5896

How ingenious! It seems that constructing composite parts and full pathway takes engineering wisdom.

CONTACT US: iGEMHUSTChina@163.com

Huazhong University of Sci. & Tech., Wuhan, China

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