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米曲霉钙调蛋白编码基因(cmdA)的克隆与核苷酸序列分析

Cloning and nucleotide sequence of the calmodulin-encoding gene (cmdA) from Aspergillus oryzae.

作者信息

Yasui K, Kitamoto K, Gomi K, Kumagai C, Ohya Y, Tamura G

机构信息

Research Institute of Brewing Resources, Co., Ltd., Tokyo, Japan.

出版信息

Biosci Biotechnol Biochem. 1995 Aug;59(8):1444-9. doi: 10.1271/bbb.59.1444.

Abstract

A cDNA and genomic gene encoding calmodulin were isolated from Aspergillus oryzae using a part of the calmodulin gene from A. nidulans as a hybridization probe. The gene was in a 3.4-kb SphI fragment and Southern-blot analysis of genomic DNA suggested the existence of a single copy of the calmodulin gene in A. oryzae. The nucleotide sequence analysis showed that the gene consists of five introns and six exons. Although the nucleotide sequence homology with that of A. nidulans was not so high (68%), the deduced amino acid sequence was 100% and 84% identical with calmodulin of A. nidulans and chicken, respectively. The cDNA encoding A. oryzae calmodulin was expressed under the control of the GAL1 promoter in the calmodulin null mutant (cmd1) of yeast, Saccharomyces cerevisiae, and could function as a calmodulin gene.

摘要

以构巢曲霉钙调蛋白基因的一部分作为杂交探针,从米曲霉中分离出编码钙调蛋白的cDNA和基因组基因。该基因位于一个3.4kb的SphI片段中,对基因组DNA的Southern印迹分析表明米曲霉中存在钙调蛋白基因的单拷贝。核苷酸序列分析表明该基因由5个内含子和6个外显子组成。虽然与构巢曲霉的核苷酸序列同源性不高(68%),但推导的氨基酸序列分别与构巢曲霉和鸡的钙调蛋白100%和84%相同。编码米曲霉钙调蛋白的cDNA在酿酒酵母钙调蛋白缺失突变体(cmd1)中受GAL1启动子控制表达,并可作为钙调蛋白基因发挥功能。

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