在小菌落阴性乳酸克鲁维酵母中编码乙酰鸟氨酸转氨酶的ARG8基因的克隆、测序及破坏。

Cloning, sequencing and disruption of the ARG8 gene encoding acetylornithine aminotransferase in the petite-negative yeast Kluyveromyces lactis.

作者信息

Janssen A, Chen X J

机构信息

Research School of Biological Sciences, Australian National University, Canberra City, Australia.

出版信息

Yeast. 1998 Feb;14(3):281-5. doi: 10.1002/(SICI)1097-0061(199802)14:3<281::AID-YEA212>3.0.CO;2-E.

DOI:10.1002/(SICI)1097-0061(199802)14:3<281::AID-YEA212>3.0.CO;2-E

PMID:9544247

Abstract

A recombinant plasmid was isolated from a Kluyveromyces lactis genomic DNA library which complements a Saccharomyces cerevisiae arg8 mutant defective in the gene encoding acetylornithine aminotransferase. The complementation activity was found to reside within a 2.0 kb DNA fragment. Nucleotide sequence analysis revealed an open reading frame able to encode a 423-residue protein sharing 68.1% and 35.0% sequence identities with the products of the ARG8 and argD genes of S. cerevisiae and Escherichia coli. That the cloned gene, KlARG8, is the functional equivalent of S. cerevisiae ARG8 was supported by a gene disruption experiment which showed that K. lactis strains carrying a deleted chromosomal copy of KlARG8 are auxotrophic for arginine.

摘要

从乳酸克鲁维酵母基因组DNA文库中分离出一种重组质粒，该质粒可互补酿酒酵母中编码乙酰鸟氨酸转氨酶的基因发生缺陷的arg8突变体。发现互补活性存在于一个2.0 kb的DNA片段中。核苷酸序列分析揭示了一个开放阅读框，其能够编码一种423个残基的蛋白质，该蛋白质与酿酒酵母和大肠杆菌的ARG8和argD基因产物的序列同一性分别为68.1%和35.0%。基因破坏实验支持克隆的基因KlARG8与酿酒酵母ARG8功能等同，该实验表明携带KlARG8染色体缺失拷贝的乳酸克鲁维酵母菌株对精氨酸营养缺陷。