Difference between revisions of "Part:BBa K1763426"
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This construct was successfully amplified using the [http://2015.igem.org/Team:UCLA/Notebook/Spider_Silk_Genetics/ICA_Oligo_Sequences post-elution primers] after ICA. The results of the amplification are shown below. | This construct was successfully amplified using the [http://2015.igem.org/Team:UCLA/Notebook/Spider_Silk_Genetics/ICA_Oligo_Sequences post-elution primers] after ICA. The results of the amplification are shown below. | ||
| − | [[File:7 13 2015 UCLA ICA 2.jpg|none|thumb|500px|'''Fig. 1''' PCR amplification of MaSp2 6 and 9 mer using post-elution primers. The expected product size for MaSp2-6 is 712 bp.]] | + | [[File:7 13 2015 UCLA ICA 2.jpg|none|thumb|500px|'''Fig. 1''' PCR amplification of MaSp2 6 and 9 mer using post-elution primers at different temperatures. The expected product size for MaSp2-6 is 712 bp.]] |
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===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K1763426 parameters</partinfo> | <partinfo>BBa_K1763426 parameters</partinfo> | ||
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Revision as of 20:26, 18 September 2015
MaSp2-6(1C3)
This is a 6-mer construct of MaSp2 constructed using Iterative Capped Assembly (ICA) protocols as outlined on the 2015 UCLA iGEM page [http://2015.igem.org/Team:UCLA/Notebook/Spider_Silk_Genetics/Protocols here]. This part is a coding sequence, with no regulatory elements present. This construct includes an N-terminal 8-his tag which can be used purify this protein using nickel based affinity purification techniques.
Biology
Native spider silk genes consist of many repeats of the MaSp genes, up to 100 repeats. The extensive repetition confers the properties of strength and elasticity that are commonly associated with spider silk threads. In order to examine the role of protein length and composition on the final properties of spider silk, we created this construct using Iterative Capped Assembly.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 652
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage
In order to express this construct as a protein, it is necessary to subclone it after a promoter and RBS. The 2015 UCLA iGEM team has made a composite part, BBa_K1764427, has placed this part under control of the T7-RBS regulatory elements.
This construct was successfully amplified using the [http://2015.igem.org/Team:UCLA/Notebook/Spider_Silk_Genetics/ICA_Oligo_Sequences post-elution primers] after ICA. The results of the amplification are shown below.
