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枯草芽孢杆菌ribC基因的分子克隆与特性分析:ribC中的一个点突变导致核黄素过量产生。

Molecular cloning and characterisation of the ribC gene from Bacillus subtilis: a point mutation in ribC results in riboflavin overproduction.

作者信息

Coquard D, Huecas M, Ott M, van Dijl J M, van Loon A P, Hohmann H P

机构信息

Biotechnology Section of the Vitamin Division (VFCB), Hoffmann-La Roche AG, Basel, Switzerland.

出版信息

Mol Gen Genet. 1997 Mar 18;254(1):81-4. doi: 10.1007/s004380050393.

Abstract

A mutation leading to roseoflavin resistance and deregulated riboflavin biosynthesis was mapped in the genome of the riboflavin-overproducing Bacillus subtilis strains RB52 and RB50 at map position 147 degrees. The chromosomal location indicates that the deregulating mutation in RB52 and RB50 is an allele of the previously identified ribC mutation. We cloned the ribC gene and found that it encodes a putative 36-kDa protein. Surprisingly, RibC has significant sequence similarity to flavin kinases and FAD synthases from various other bacterial species. By comparing the deduced amino acid sequence of RibC from the wild-type parent strain of RB50 with the RibC sequence from the riboflavin-overexpressing RB50 mutant we identified a point mutation that resulted in a Gly to Ser exchange in the C-terminal region of the product.

摘要

在核黄素高产枯草芽孢杆菌菌株RB52和RB50的基因组中,位于图谱位置147°处定位到一个导致玫瑰黄素抗性和核黄素生物合成失调的突变。染色体定位表明,RB52和RB50中的失调突变是先前鉴定的ribC突变的一个等位基因。我们克隆了ribC基因,发现它编码一种推定的36 kDa蛋白。令人惊讶的是,RibC与来自其他各种细菌物种的黄素激酶和FAD合酶具有显著的序列相似性。通过比较RB50野生型亲本菌株的RibC推导氨基酸序列与核黄素过表达的RB50突变体的RibC序列,我们鉴定出一个点突变,该突变导致产物C末端区域的甘氨酸到丝氨酸交换。

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