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多形汉逊酵母硝酸还原酶脱辅基酶编码基因YNR1的克隆与破坏

Cloning and disruption of the YNR1 gene encoding the nitrate reductase apoenzyme of the yeast Hansenula polymorpha.

作者信息

Avila J, Pérez M D, Brito N, González C, Siverio J M

机构信息

Departamento de Bioquímica y Biología Molecular, Universidad de La Laguna, Tenerife, Canarias, Spain.

出版信息

FEBS Lett. 1995 Jun 12;366(2-3):137-42. doi: 10.1016/0014-5793(95)00511-7.

DOI:10.1016/0014-5793(95)00511-7
PMID:7789531
Abstract

The nitrate reductase gene (YNR1) from the yeast H. polymorpha was isolated from a lambda EMBL3 genomic DNA library. As probe a 350 bp DNA fragment synthesized by PCR from H. polymorpha cDNA was used. By DNA sequencing an ORF of 2,577 bp was found. The predicted protein has 859 amino acids and presents high identity with nitrate reductases from other organisms. Chromosomal disruption of YNR1 causes inability to grow in nitrate. Northern blot analysis showed that YNR1 expression is induced by nitrate and repressed by ammonium.

摘要

从多形汉逊酵母（H. polymorpha）的λEMBL3基因组DNA文库中分离出硝酸还原酶基因（YNR1）。使用通过PCR从多形汉逊酵母cDNA合成的350 bp DNA片段作为探针。通过DNA测序发现了一个2577 bp的开放阅读框（ORF）。预测的蛋白质有859个氨基酸，与其他生物体的硝酸还原酶具有高度同源性。YNR1的染色体破坏导致无法在硝酸盐中生长。Northern印迹分析表明，YNR1的表达受硝酸盐诱导，受铵抑制。

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