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耐热芽孢杆菌E2菌株纤维素酶基因克隆的研究 被引量:6

Molecular Cloning of Cellulase Gene from the Bacillus sp,Strain E2
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摘要 用质位pBR322作载体,大肠杆菌E.coliDH5αF'作受体,克隆到耐热芽孢杆菌E2菌株的羧甲基纤维素酶(CMCase)基因。重组质粒pBG3从产生CMCase的转化子中分离得到。克隆到的CMCare基因位于4.0kbHindIII片段上。功能状态CMCase基因被亚克隆到2.4kbDNA片段上。同位素杂交结果显示出克隆到的CMCase基因与供体菌染色体DNA的亲合性。重组质粒pBG3所表达的CMCase的最适作用pH为6.5,最适作用温度为55℃,不作用于任何实验过的天然纤维素。含重组质粒pBG3的转化子产生的纤维素酶在细胞中的分布为:胞内占82.9%,胞外占7.9%,质壁空间(periplasmicspace)占9.2%。 The CMCase gene from the Bacillus sp. strain E2 was cloned into E,coli DH5αF'using plasmid pBR322 as a vector. Recombinant plasmid pBG3 was isolated from transformants producing CMCase,and the cloned CMCasc gene was found to be in a 4.0kb HindIII fragment. The CMCase gene was subcloned in a functional state on a 2.4kb DNA fragment.The optimum pH and temperature for activity of CMCase encoded by pBG3 were pH 6.5 and 55℃ respectively.The CMCase could not hydrolyze any natural cellulose examined.Eighty-three percent of the CMCase was detected in the inner cellular fraction,9%and 8%were in the periplasmic space and medium respectively.Result of Southern hybridization indicates that the cloned DNA fragment was derived from Bacillus sp.strain E2 chromosomal DNA. Received May 11,1994.This Project Supported by National Natural Science Foundation of China
出处 《Acta Genetica Sinica》 SCIE CAS CSCD 1995年第4期322-328,共7页
基金 国家自然科学基金
关键词 芽孢杆菌 E2菌株 基纤维素酶 基因克隆 Bacillus sp.strain E2,CMCase,Gene cloning
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