Part:BBa_K1884012
RBCS2 Promoter+Hygromycin B+RBCS2 Terminator
We provided Hygromycin B resistance under the RBCS2 promoter(BBa_K1884010)
Biology
RBCS2 is a promoter of the small subunit of ribulose-1, 5-diphosphate carboxylase, encoded by the nuclear gene and is an inducible promoter for strong light-induced expression.
Hygromycin is active against both prokaryotic and eukaryotic cells. It acts by inhibiting polypeptide synthesis. It stabilizes the tRNA-ribosomal acceptor site, thereby inhibiting translocation. .
RBCS2 terminator is Chlamydomonas reinhardtii RuBisCO small subunit 2 terminator which obtained from the pOpt_mVenus_Hyg expression plasmid provided by Chlamydomonas Resource Center.(Fig 1)
Usage
For testing the efficiency of Hygromycin B, we formulated six gradient concentrations of Hygromycin B solution and add 200ul each into six different beakerflask which contain 200ml LB solid medium, then we put the competent cell into six different LB plate. there is a photograph to show several monoclonal colony growing on a LB plate which has the best selected abilty of Hygromycin B.(Fig 2)
Conbined with another resistant device(BBa_K1884011), we are able to screen the green algae which has paromoycin resistance and Hygromycin B resistance. we added 200ul paromoycin solution and hygromycin B solution into 200ml TAP solid medium, then we put the green algae without cytoderm into TAP plate. there is a photograph to show several colony growing on a TAP plate which has the best selected efficiency of paromoycin and Hygromycin B.(Fig 3)
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 650
Illegal NgoMIV site found at 832 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1105
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