摘要
采用PCR技术 ,从GFPmut2中扩增得到三位点突变的报告基因gfpS65T、V68L、S72A片段 ,并将它和肺炎克氏杆菌(Klebsiellapneumoniae (Schr eter)Trevisan)M5a1的固氮酶结构基因nifH的启动子和其起始密码子相融合 ,获得nifH_gfp表达载体pMGFP2 ;再在pMGFP2上插入卡那霉素抗性基因 ,获得可在日勾维肠杆菌 (Enterobactergergoviae)5 7_7表达nifH_gfp的表达载体pMGFP2 .1。研究了此表达载体经转化E .gergoviae 5 7_7后 ,NH+ 4和氧对E .gergoviae5 7_7中nifH_gfp表达的影响。
A nif H_ gfp expression vector pMGFP2 was constructed by fusing the 725 bp PCR amplified triple mutated green fluorescent protein ( GFP ) gene ( gfp S65T,V68L,S72A ) fragment to the nif H promoter and its start codon which was from Klebsiella pneumoniae (Schreter) Trevisan M5a1. A kanamycin cassette was inserted into Pst Ⅰ site of pMGFP2, obtaining the expressing vector pMGFP2.1 which can be used for the studying of nif H_ gfp expression in Enterobacter gergoviae 57_7. It was then transformed into E.gergoviae 57_7 and the effects of NH + 4 and oxygen on the expression of nif H_ gfp in E. gergoviae 57_7 were studied.
基金
国家自然科学基金资助项目! (39570 0 65)
国家高技术研究发展计划"863"资助项目!(1 0 1_0 3_0 4_0 2 )&&
