Difference between revisions of "Part:BBa K1884012"

Revision as of 20:32, 16 October 2016

RBCS2 Promoter+Hygromycin B+RBCS2 Terminator

We provided Hygromycin B resistance under the RBCS2 promoter(BBa_K1884010)

Biology

RBCS2 promoter

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.

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 1)

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 2)

Sequence and Features