Genetic transformation of marine cyanobacterium Synechococcus sp.CC9311(Cyanophyceae) by electroporation  

作　　者：陈华新 林瀚智 姜鹏 李富超 秦松 

机构地区：[1]Key Laboratory of Experimental Marine Biology,Institute of Oceanology,Chinese Academy of Sciences [2]Graduate University of Chinese Academy of Sciences [3]Yantai Institute of Coastal Zone Research,Chinese Academy of Sciences

出　　处：《Chinese Journal of Oceanology and Limnology》2013年第2期416-420,共5页中国海洋湖沼学报（英文版）

基　　金：Supported by the Key Innovation Project of Institute of Oceanology,Chinese Academy of Sciences(No.2009-2);the Natural Science Foundation of Shandong Province(No.2009ZRB02542);the Foundation of Key Laboratory of Marine Bioactive Substance and Modern Analytical Techniques,SOA(No.MBSMAT-2010-03);the National Natural Science Foundation of China(No.41276164);the Natural Science Foundation of Jiangsu Province(No.BK2012650)

摘　　要：Synechococcus sp.CC9311 is a marine cyanobacterium characterized by type IV chromatic acclimation(CA).A genetic transformation system was developed as a first step to elucidate the molecular mechanism of CA.The results show that Synechococcus sp.CC9311 cells were sensitive to four commonly used antibiotics:ampicillin,kanamycin,spectinomycin,and chloramphenicol.An integrative plasmid to disrupt the putative phycoerythrin lyase gene mpeV,using a kanamycin resistance gene as selectable marker,was constructed by recombinant polymerase chain reaction.The plasmid was then transformed into Synechococcus sp.CC9311 via electroporation.High transformation efficiency was achieved at a field strength of 2 kV/cm.DNA analysis showed that mpeV was fully disrupted following challenge of the transformants with a high concentration of kanamycin.In addition,the transformants that displayed poor growth on agar SN medium could be successfully plated on agarose SN medium.Synechococcus sp. CC9311 is a marine cyanobacterium characterized by type IV chromatic acclimation （CA）. A genetic transformation system was developed as a first step to elucidate the molecular mechanism of CA. The results show that Synechococcus sp. CC9311 cells were sensitive to four commonly used antibiotics： ampicillin, kanamycin, spectinomycin, and chloramphenicol. An integrative plasmid to disrupt the putative phycoerythrin lyase gene mpeV, using a kanamycin resistance gene as selectable marker, was constructed by recombinant polymerase chain reaction. The plasmid was then transformed into Synechococcus sp. CC9311 via electroporation. High transformation efficiency was achieved at a field strength of 2 kV/cm. DNA analysis showed that mpeV was fully disrupted following challenge of the transformants with a high concentration of kanamycin. In addition, the transformants that displayed poor growth on agar SN medium could be successfully plated on agarose SN medium.

关 键 词：chromatic acclimation ELECTROPORATION genetic transformation Synechococcus sp. CC9311 

分 类 号：Q943[生物学—植物学]