【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 7₇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 7₇后 ,NH+ 4和氧对E .gergoviae5 7₇中nifH_gfp表达的影响。更多还原

【Abstract】 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 （Schreter） 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 57₇. It was then transformed into E.gergoviae 57₇ and the effects of NH + 4 and oxygen on the expression of nif H gfp in E. gergoviae 57₇ were studied.更多还原