巴西固氮螺菌中吸氢酶基因同源性的分子检测  

MOLECULAR DETECTION OF HOMOLOGY OF HYDROGEN UPTAKING GENE IN Azospirillu brasilense

作　　者：罗永华[1] 郭俊[1] 

机构地区：[1]广东省微生物研究所,广东广州510070

出　　处：《微生物学杂志》2002年第3期12-13,34,共3页Journal of Microbiology

摘　　要：通过TTC(2 ,3,5 氯化三苯基氮唑 )实验筛选出 12株能产生吸氢酶蛋白 (Hup+ )的巴西固氮螺菌 (Azospir rilumbrasilense)菌株。用Qiagen柱分离提纯含豌豆根瘤菌的hup基因片段的质粒 pHVT10 9和pHVT115 ,并用地高辛标记法标记pHVT115 ,与 12株Hup+ 巴西固氮螺菌的总DNA进行斑点杂交 ,结果显示pHVT115所含的吸氢酶基因 (hup基因 )片段与巴西固氮螺菌的大部分菌株的hup基因同源性不强。这一结果表明hup基因在固氮生物中存在着遗传多样性 ,异源hup基因探针不一定都适宜于探测hup基因。Azospirillum brasilense strains capable of producing hydrogen uptaking enzyme protein (Hup\++) were screened through TTC (2,3,5 triphenyl tetrazolium chloride) experiment. Plasmids pHVT109 and pHVT115 containing fragment of hup gene of Rhizobium leguminosarum isolated and purified with Qiagen column was obtained. As a probe pHVT115 was labeled with digoxigenin Dutp and dot hybridized with total DNA of 12 Hup\++ A. brasilense strains. The results showed that there was only weak homology between hup genes of most of A. brasilense strains and hup gene contained in pHVT 115. It also revealed that hup gene existed genetic diversity in nitrogen fixing organisms. And heterological genetic probe could not be always suitable for detecting hup genes.\;

关 键 词：巴西固氮螺菌 吸氢酶基因 斑点杂交 

分 类 号：S154.32[农业科学—土壤学]