Difference between revisions of "Part:BBa K1958004"
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| + | ==Usage and Biology== | ||
This part HyaD encodes for the fourth subunit of hydrogenase 1 in E.coli. | This part HyaD encodes for the fourth subunit of hydrogenase 1 in E.coli. | ||
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Figure 1. The Hya operon on the genome of E.coli. | Figure 1. The Hya operon on the genome of E.coli. | ||
| − | == Results == | + | ==Results== |
First the recombinant plasmid with genes encoding E.coli hydrogenase 1 was constructed. This enzyme is encoded by hya operon where lie in all six genes named hyaABCDEF (BBa_K1958001, BBa_K1958002, BBa_K1958003, BBa_K1958004, BBa_K1958000, BBa_K1958006) on the genome of E.coli. The sequence was PCR amplified according to the sequence from our vessel E.coli strain BL21(DE3). We have completed the plasmid pET28a with hya cluster promoted by a T7 promoter. The following figure shows the result of enzyme digestion assay of recombinant plasmid in which our target gene cluster displayed a band around 5.5kb, which indicated successful hydrogenase expression. | First the recombinant plasmid with genes encoding E.coli hydrogenase 1 was constructed. This enzyme is encoded by hya operon where lie in all six genes named hyaABCDEF (BBa_K1958001, BBa_K1958002, BBa_K1958003, BBa_K1958004, BBa_K1958000, BBa_K1958006) on the genome of E.coli. The sequence was PCR amplified according to the sequence from our vessel E.coli strain BL21(DE3). We have completed the plasmid pET28a with hya cluster promoted by a T7 promoter. The following figure shows the result of enzyme digestion assay of recombinant plasmid in which our target gene cluster displayed a band around 5.5kb, which indicated successful hydrogenase expression. | ||
Revision as of 14:52, 22 October 2016
HyaD -> E. coli
HyaD is a subunit of hydrogenase 1 (Hyd-1) from Escherichia coli genome. The hya operon, which contains gene for the two HYD1 structual subunits and four additional genes (HyaA-F) was mapped at 22min on the E.coli chromosome. And hyaD is the fourth gene of hya operon, sharing the first few base pairs with hyaC and the last few base pairs with hyaE. HyaD is the specific protease required for large subunit maturation-terminal processing. HyaD is a hydrogenase maturase.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]