Difference between revisions of "Part:BBa K1897000:Design"
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===Design Notes=== | ===Design Notes=== | ||
− | We have included a hexa-histidine tag at the end of the coding sequence as to facilitate protein purification using Immobilised Metal Affinity Chromatography. However, this is only the coding sequence and therefore a relevant promoter, ribosome binding site and terminator would be required for expression (see | + | We have included a hexa-histidine tag at the end of the coding sequence as to facilitate protein purification using Immobilised Metal Affinity Chromatography. However, this is only the coding sequence and therefore a relevant promoter, ribosome binding site and terminator would be required for expression (see BBa_K1897001 for the HasA construct that is ready for expression). This construct can be used by others who wish to change the control of the expression of this gene by adding their own promoters. |
===Source=== | ===Source=== |
Revision as of 16:18, 9 October 2016
HasA hemophore coding sequence
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 442
Design Notes
We have included a hexa-histidine tag at the end of the coding sequence as to facilitate protein purification using Immobilised Metal Affinity Chromatography. However, this is only the coding sequence and therefore a relevant promoter, ribosome binding site and terminator would be required for expression (see BBa_K1897001 for the HasA construct that is ready for expression). This construct can be used by others who wish to change the control of the expression of this gene by adding their own promoters.
Source
The HasA gene is originally found in the Serratia marcescens. We obtained the gene sequence from the National Center for Biotechnology Information (NCBI) database (http://www.ncbi.nlm.nih.gov/nuccore/X81195.2) and synthesised the gene with the hexa histadine tag.
References
Serratia marcescens hasA, has B, hasD, hasE, hasI, hasR and hasS genes. Retrieved May 2016 from http://www.ncbi.nlm.nih.gov/nuccore/X81195.2
Cescau, S., Cwerman, H., Letoffe, S., Delepelaire, P., Wandersman, C., & Biville, F. (2007). Heme acquisition by hemophores. Biometals, 20(3-4), 603-613.