Construction of a Novel Shuttle Vector for Use in <Emphasis Type="Italic">Gluconobacter oxydans</Emphasis> |
| |
Authors: | Lin Zhang Jinping Lin Yushu Ma Dongzhi Wei Ming Sun |
| |
Institution: | (1) State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, China;(2) School of Mechanical and Power Engineering, East China University of Science and Technology, Shanghai, 200237, China; |
| |
Abstract: | 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. |
| |
Keywords: | |
本文献已被 SpringerLink 等数据库收录! |
|