Difference between revisions of "Part:BBa K3332020"
| Line 12: | Line 12: | ||
1. Agarose Gel Electrophoresis | 1. Agarose Gel Electrophoresis | ||
After receiving the synthesized DNA, restriction digestion was done to certify that the plasmid was correct, and the experimental results were shown in figure 1. | After receiving the synthesized DNA, restriction digestion was done to certify that the plasmid was correct, and the experimental results were shown in figure 1. | ||
| − | <table><tr><th>[[File:T--XMU-China2020--BBa K3332020 2. | + | <table><tr><th>[[File:T--XMU-China2020--BBa K3332020 2.jpg|thumb|300px|Fig.1 The result of plasmid cut with enzyme ''EcoR''I and ''PstI''. Plasmid: pSB1C3..]]</th><th></table> |
<!-- --> | <!-- --> | ||
Revision as of 03:06, 25 October 2020
phnE1
Subunit of phosphonate ABC transporter, permease protein phnE, from S.meliloti 1021.Use K823004 to construct a new part that can transport glyphosate to cytoplasm.
Biology
Phn system is a gene cluster for organophosphorus transport and degradation in many microorganisms. Sinorhizobium meliloti 1021 use PhnE1 and PhnE2 to construct the permease protein of ABC transporter, which includes PhnE1, PhnE2, PhnC and PhnD proteins, this transporter can transport glyphosate to cytoplasm. The phnE1 gene encodes a subunit of permease protein which can transport glyphosate to cytoplasm.
Usage
We ligased the strong promoter (BBa_J23100) and the parts (RBS-phnE1-Terminator) on the expression vector pSB1C3 by standard assembly. Then the ligation mixture was transformed into E. coli DH5α & E. coli BL21(DE3), enabled the E. coli to express PhnE1 protein.
Characterization
1. Agarose Gel Electrophoresis After receiving the synthesized DNA, restriction digestion was done to certify that the plasmid was correct, and the experimental results were shown in figure 1.
File:T--XMU-China2020--BBa K3332020 2.jpg Fig.1 The result of plasmid cut with enzyme EcoRI and PstI. Plasmid: pSB1C3.. |
|---|
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]