文献类型：期刊文献

英文题名：Construction of a Novel Shuttle Vector for Use in Gluconobacter oxydans

作者：Zhang, Lin[1];Lin, Jinping[1];Ma, Yushu[1];Wei, Dongzhi[1];Sun, Ming[2]

机构：[1]E China Univ Sci & Technol, New World Inst Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]E China Univ Sci & Technol, Sch Mech & Power Engn, Shanghai 200237, Peoples R China

年份：2010

卷号：46

期号：3

起止页码：227

外文期刊名：MOLECULAR BIOTECHNOLOGY

收录：;EI(收录号：20104313322617);WOS:【SCI-EXPANDED(收录号:WOS:000282272500003)】；

基金：This work financially supported by the National Natural Science Foundation of China (Grant No. 20976053/B060804) and the National Special Fund for State Key Laboratory of Bioreactor Engineering (Grant No. 2060204).

语种：英文

外文关键词：Shuttle vector; Gluconobacter oxydans; Cryptic plasmid; Expression

摘要：A shuttle vector pZL1 which can replicate both in Gluconobacter oxydans and Escherichia coli was constructed based on G. oxydans DSM2003 cryptic plasmid pGOX3, a homology of G. oxydans 621H pGOX3, and E. coli cloning vector pUC18. It was found to be stably maintained in G. oxydans during the serial subcultures in the absence of antibiotic pressure for 144 h. With pGOX3 as the reference sample, the relative copy number of pZL1 in G. oxydans is 13 determined by real-time fluorescence quantitative PCR (qPCR). The copy number of pZL1 is much higher than pBBR1MCS5 in E. coli. The vector pZL1 contains six commonly used restriction endonuclease sites, HindIII, SalI, XbaI, BamHI, SmaI, KpnI, and SacI, and is easy to manipulate in molecular biology experiments. The shuttle vector was used to express a reporter protein wasabi successfully in G. oxydans DSM2003 under the control of the tufB promoter.