Difference between revisions of "Part:BBa K3888011:Design"
(→Figure 1: The Oxygen-tolerant Hydrogenase introduction plasmid constructed, which has a length of 8261bp.) |
|||
| Line 3: | Line 3: | ||
<partinfo>BBa_K3888011 short</partinfo> | <partinfo>BBa_K3888011 short</partinfo> | ||
[[File:pET28-pTac-lacO-hydrogenase .png]] | [[File:pET28-pTac-lacO-hydrogenase .png]] | ||
| − | ==Figure 1: The Oxygen-tolerant Hydrogenase introduction plasmid constructed, which has a length of 8261bp.== | + | ===Figure 1: The Oxygen-tolerant Hydrogenase introduction plasmid constructed, which has a length of 8261bp.=== |
Two genes, HoxG1 and HoxK1, which are the main components of the whole introduction plasmid. HoxG1 encodes the large subunit of the oxygen-tolerant hydrogenase and HoxK1 encodes the small subunit of the oxygen-tolerant hydrogenase.<br>The position of two genes are located from 5094bp to 6926bp and from 7137bp to 8102bp respectively. <br> | Two genes, HoxG1 and HoxK1, which are the main components of the whole introduction plasmid. HoxG1 encodes the large subunit of the oxygen-tolerant hydrogenase and HoxK1 encodes the small subunit of the oxygen-tolerant hydrogenase.<br>The position of two genes are located from 5094bp to 6926bp and from 7137bp to 8102bp respectively. <br> | ||
The two genes are promoted by pTac, a promoter. pTac is controlled by a lac operator with a LacI upstream. IPTG will induce lacI repressors to change its conformation and prevent it from combining with lac operator thus starts the transcription of genes, which are HoxG1 and HoxK1. | The two genes are promoted by pTac, a promoter. pTac is controlled by a lac operator with a LacI upstream. IPTG will induce lacI repressors to change its conformation and prevent it from combining with lac operator thus starts the transcription of genes, which are HoxG1 and HoxK1. | ||
Revision as of 13:02, 20 October 2021
[Ni-Fe] Hydrogenase Core gene HoxG1 and HoxK1
Figure 1: The Oxygen-tolerant Hydrogenase introduction plasmid constructed, which has a length of 8261bp.
Two genes, HoxG1 and HoxK1, which are the main components of the whole introduction plasmid. HoxG1 encodes the large subunit of the oxygen-tolerant hydrogenase and HoxK1 encodes the small subunit of the oxygen-tolerant hydrogenase.
The position of two genes are located from 5094bp to 6926bp and from 7137bp to 8102bp respectively.
The two genes are promoted by pTac, a promoter. pTac is controlled by a lac operator with a LacI upstream. IPTG will induce lacI repressors to change its conformation and prevent it from combining with lac operator thus starts the transcription of genes, which are HoxG1 and HoxK1.
There is an RBS located between 5068bp and 5087bp. We chose RBS:B0032m as our ribosome binding site. It's a wild-type (also referred to as the natural type).
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 363
Illegal XbaI site found at 465
Illegal SpeI site found at 531 - 12INCOMPATIBLE WITH RFC[12]Illegal SpeI site found at 531
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 678
Illegal BamHI site found at 1593
Illegal BamHI site found at 2932
Illegal XhoI site found at 1915 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 363
Illegal XbaI site found at 465
Illegal SpeI site found at 531 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 363
Illegal XbaI site found at 465
Illegal SpeI site found at 531
Illegal AgeI site found at 1809 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
Change signal sequence of HoxK1
Source
Hydrogenovibrio marinus MH-110
References
1. Kim, Jaoon YH, Byung Hoon Jo, and Hyung Joon Cha. "Production of biohydrogen by heterologous expression of oxygen-tolerant Hydrogenovibrio marinus [NiFe]-hydrogenase in Escherichia coli." Journal of biotechnology 155.3 (2011): 312-319.