摘要
利用三亲接合的方法将含有 ntrB 基因部分序列的自杀载体导入固氮斯氏假单胞菌(Pseudomonas stutzeri)A1501中,获得了 ntrB 基因的非极性突变株(AdnB)。RT-PCR实验证明,AdnB 菌株中与 ntrB 连锁的下游基因 ntrC 能正常转录;ntrB 基因的非极性突变导致细胞完全丧失固氮能力,但其突变株能在含 NH_4^+的基本培养基 K 中正常生长;在以硝酸盐为唯一氮源时,AdnB 比野生型 A1501的细胞生长延迟约14～18 h,认为 ntrB 基因与硝酸盐的氮代谢有关。功能互补实验结果表明,AdnB 的功能互补菌株能完全恢复细胞的固氮作用和对硝酸盐的利用。
The suicide vector containing partial gene sequence of ntrB was constructed and introduced into the wild type strain A1501 of Pseudomonas stutzeri by triparental conjugation to generate the ntrB mutant （AdnB）. The RT-PCR experiment showed that ntrC gene could normally transcript in the AdnB strain. Disruption of ntrB gene in the Pseudomonas stutzeri strain A1501 caused the cells to lost their nitrogen fixation ability, but the AdnB strain could grow normally in the minimal medium K with NH4^＋.With nitrate as a single nitrogen source, the growth of the AdnB strain was delayed 14-18h compared with the wild type of A1501 strain, suggesting that ntrB gene is relevant to nitrate metabolism. The results of the function complemental experiment shows that AdnBComp can fully resume the cells＇ function of nitrogen fixation and the utilization of nitrate.
出处
《高技术通讯》
EI
CAS
CSCD
北大核心
2008年第2期211-215,共5页
Chinese High Technology Letters
基金
863计划(2001AA214021)
973计划(2001CB108904)资助项目
