β－淀粉酶基因在黄单胞菌中的克隆及表达  

CLONING AND EXPRESSION OF β-AMYLASE GENE IN Xanthomonas campestris

作　　者：杨建华[1] 李时岩 孟淑芳[1] 牛淑敏[1] 王树荣[1] 王金华[1] 朱峰[1] 宋诚[1] 

机构地区：[1]南开大学微生物系,河南师范大学生物系

出　　处：《南开大学学报（自然科学版）》1996年第1期69-73,共5页Acta Scientiarum Naturalium Universitatis Nankaiensis

摘　　要：广泛寄主范围载体ｐＫＴ２１０可在辅助质粒ｐＲＫ２０１３的协助下转移进入黄单胞菌（革兰氏阴性）中并能稳定存在；来源于枯草杆菌的β－淀粉酶基因与载体ｐＫＴ２１０形成重组质粒ｐＹＬ１，并以其转化大肠杆菌；通过三亲本接合将ｐＹＬ１引入带有利福平抗性的黄单胞菌ＮＫ０１－Ｒ中，通过抗性选择接合子，并测定接合子的淀粉酶水解能力及产胶能力．本文获得一株黄原胶产量比出发菌株提高２０％，且淀粉酶水解能力显著提高的工程菌株．Broad-host range cloning vector pKT210 could be mobilized into Gram-negative bacteria X. campestris by helping of helper plasmid pRK2013. β-amylase gene origined from B.subtilis was ligated with vector pKT210 into recombinant pYL1 and then was transfered into E. colt LE392. Transfermants were selected by indine vapour. PYL1 was introduced into Rif-resistence X. campestris NK-of by triparental conjugation. Exoconjugants were screened for R1f-,Cm- and Sm-resistance marker, and their hydrolysis ability, xanthan gum producing ability were measured. A genetic engineering strain with highly improved hydrolysis ability and more than 30% xanthan gum production was obtained.

关 键 词：黄单胞菌 重组质粒 Β-淀粉酶 基因表达 

分 类 号：Q556[生物学—生物化学]