CLONING OF A NOVEL CRYIC-TYPE GENE FROM A STRAIN OF BACILLUS-THURINGIENSIS SUBSP GALLERIAE
CLONING OF A NOVEL CRYIC-TYPE GENE FROM A STRAIN OF BACILLUS-THURINGIENSIS SUBSP GALLERIAE
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DOI：
10.1128/aem.59.4.1131-1137.1993
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发表时间：
1993-04-01
影响因子：
4.4
通讯作者：
YAMAMOTO, T
中科院分区：
文献类型：
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作者：
KALMAN, S;KIEHNE, KL;YAMAMOTO, T
从苏云金芽孢杆菌（Bacillus thuringiensis subsp.）galleriae。一种新的聚合酶链反应（PCR）技术与一套几个寡核苷酸引物对特异性的cryIC基因被用来筛选一些B。苏云金杆菌菌株PCR扩增出B的几个DNA片段，范围从100 bp到1 kb。含有cryIC基因的苏云金杆菌菌株。从苏云金芽孢杆菌苏云金亚种（Bacillus thuringiensis subsp. galleriae HD 29 DNA片段与其他含cryIC的菌株DNA片段不同，表明菌株HD 29含有一个新的cryIC型基因。为了分离与cryIC同源的晶体基因，用从苏云金芽孢杆菌（Bacillus thuringiensis subsp.）相泽HD229。从与cryIC探针强烈杂交的噬菌体中分离出推定的毒素基因。推测的毒素DNA序列的翻译揭示了1，176个氨基酸的开放阅读框架，其预测分子量为132.8 kDa。毒素基因序列与其他cry基因序列的比较表明，该基因是cryIC的一个亚类。我们建议将该基因命名为cryIC（B）。在大肠杆菌中，cryIC（B）基因产生对甜菜夜蛾和粉纹夜蛾有毒性的约130 kDa的蛋白。
A novel cryIC-type gene was isolated from a strain of Bacillus thuringiensis subsp. galleriae. A new polymerase chain reaction (PCR) technique with a set of several oligonucleotide primer pairs specific to the cryIC gene was used to screen a number of B. thuringiensis strains. PCR amplified several DNA fragments ranging from 100 bp to 1 kb for B. thuringiensis strains containing a cryIC gene. PCR fragments amplified from the Bacillus thuringiensis subsp. galleriae HD29 DNA differed from the fragments amplified from other cryIC-containing strains, indicating strain HD29 contained a novel cryIC-type gene. To isolate crystal genes homologous to cryIC, an HD29 gene library was probed with a 984-bp fragment of the amino-terminal coding region of the cryIC gene cloned from Bacillus thuringiensis subsp. aizawai HD229. A putative toxin gene was isolated from a phage that hybridized strongly to the cryIC probe. Translation of the putative toxin DNA sequence revealed an open reading frame of 1,176 amino acids whose predicted molecular mass was 132.8 kDa. Comparisons of the toxin gene sequence with sequences of other cry genes indicated that this gene is a subclass of cryIC. We propose to designate this gene cryIC(b). In Escherichia coli, the cryIC(b) gene produced a protein of approximately 130 kDa toxic to Spodoptera exigua and Trichoplusia ni.