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苏云金芽孢杆菌以色列变种HD133的cryIA(b)基因在大肠杆菌中表达的完整核苷酸序列及假定启动子区域的鉴定

Complete nucleotide sequence and identification of a putative promoter region for the expression in Escherichia coli of the cryIA(b) gene from Bacillus thuringiensis var. aizawai HD133.

作者信息

Chak K F, Jen J C

机构信息

Institute of Biochemistry, National Yang-Ming Medical College, Taipei, Taiwan, Republic of China.

出版信息

Proc Natl Sci Counc Repub China B. 1993 Jan;17(1):7-14.

PMID:8234543
Abstract

The sequence of a cry gene from Bacillus thuringiensis var. aizawai HD133 was determined. This cry gene encodes a protein of 1155 amino acids, the molecular weight of which is 130622 Da. When the nucleotide sequence of this cry gene was compared with the nucleotide sequence from B.t. var. aizawai IPL7 and var. berliner 1715, only five nucleotide changes were found. Therefore, this cry gene should be grouped into the cryIA(b) gene type. A 72 nucleotide sequence upstream of the open reading frame of the cry gene was enough to conduct the transcription of the cry gene in E. coli. Using a promoter probe vector system, it was demonstrated that any DNA fragment containing this 72 nt sequence exhibited promoter activity in vivo. It is likely that a putative promoter sequence may be present within this 72 nucleotides region for the expression of this cry gene in E. coli.

摘要

测定了苏云金芽孢杆菌库斯塔克亚种HD133的cry基因序列。该cry基因编码一个由1155个氨基酸组成的蛋白质,其分子量为130622道尔顿。当将该cry基因的核苷酸序列与来自苏云金芽孢杆菌库斯塔克亚种IPL7和库斯塔克亚种1715的核苷酸序列进行比较时,仅发现了五个核苷酸变化。因此,该cry基因应归类为cryIA(b)基因类型。cry基因开放阅读框上游72个核苷酸的序列足以在大肠杆菌中指导cry基因的转录。使用启动子探针载体系统证明,任何包含该72 nt序列的DNA片段在体内均表现出启动子活性。在这72个核苷酸区域内可能存在一个推定的启动子序列,用于该cry基因在大肠杆菌中的表达。

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