慢生型大豆根瘤菌染色体的16kb EcoRⅠDNA片段的亚克隆测序分析  

Subcloning and sequencing analysis of a 16 kb Eco RⅠ DNA fragment of the chromosome of Bradyrhizobium japonicum

作　　者：唐咸来[1] 武波[2] 柏学亮[2] 唐东阶[2] 吕安国[1] 唐纪良[2] 马庆生[2] 

机构地区：[1]中国科学院沈阳应用生态研究所,辽宁沈阳110015 [2]广西大学农业部农业分子遗传重点开放实验室,广西南宁530005

出　　处：《广西大学学报（自然科学版）》2001年第2期150-155,158,共7页Journal of Guangxi University（Natural Science Edition）

基　　金：国家高技术"863"计划! ( 863 1 0 1 0 3 0 3 0 1 )

摘　　要：亚克隆测序分析发现 ,在 Bradyrhizobium japonicum的 GX2 0 1菌株染色体的 1 6kb Eco R DNA区段上含有一个与 put A基因同源的基因的 ORF30 54( Open Reading Frame) .该基因 ORF长 30 54bp,在核苷酸水平上与已报道的 put A基因有 94 %一致性 .其推断性的编码产物在氨基酸水平上与 put A编码产物脯氨酸脱氢酶 ( Pro DH)有 95%一致性 .利用 Tn5gus A5诱变的方法获得了该同源基因的标记置换突变体 ,该突变体在液体培养基 ( YMA)The results of the subcloning and sequencing analysis showed that there is a perfect ORF3054 (Open Reading Frame) homologus with the put A gene in the 16kb Eco RⅠ DNA fragment cloned from the chromosome of Bradyrhizobium japonicum GX201,the length of which was 3054 base pairs. It shared 94% identity with the DNA of the put A gene at the nucleotide level, and its putative product protein showed 95% identity with the proline dehydrogenase (ProDH) encoded by put A gene at amino acid level. When cultured in the liquid medium YMA,the growth ratio of the markerexchanger mutant of this homologous gene was lower than that of the wildtype strain.

关 键 词：B.Japonicum PutA基因 同源性 慢生型大豆根瘤菌 染色体 亚克隆 测序 

分 类 号：S154.381[农业科学—土壤学] Q785[农业科学—农业基础科学]