构巢曲霉腺苷酰硫酸激酶基因的克隆与特性分析

Cloning and characterisation of the adenosyl phosphosulphate kinase gene from Aspergillus nidulans.

作者信息

Clarke D L, Newbert R W, Turner G

机构信息

Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield, South Yorkshire, S10 2UH, UK.

出版信息

Curr Genet. 1997 Dec;32(6):408-12. doi: 10.1007/s002940050295.

DOI:10.1007/s002940050295

PMID:9388296

Abstract

The sD gene of Aspergillus nidulans has been cloned by heterologous screening of rationally selected cosmids. Co-transformation of the sD50 mutant JMP1 confirmed the presence of a functional gene. Sequence analysis determined this gene to be 680 bp in length, containing a 59-bp intron and encoding a protein of 206 amino acids. A protein-sequence comparison revealed a similarity to the C-terminal region of ATP sulphurylase, the sC gene product. Further sequence comparison revealed differences in a consensus sequence ATP-binding motif, indicating non-functionality of the APS kinase-like domain of ATP sulphurylase, and confirms sD as the gene encoding APS kinase in A. nidulans.

摘要

通过对合理选择的黏粒进行异源筛选，克隆了构巢曲霉的sD基因。对sD50突变体JMP1进行共转化，证实了功能基因的存在。序列分析确定该基因长度为680 bp，包含一个59 bp的内含子，编码一个206个氨基酸的蛋白质。蛋白质序列比较显示与ATP硫酸化酶（sC基因产物）的C末端区域相似。进一步的序列比较揭示了ATP结合基序共有序列的差异，表明ATP硫酸化酶的APS激酶样结构域无功能，并证实sD是构巢曲霉中编码APS激酶的基因。