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首页> 外文期刊>Life sciences >cDNA cloning and expression of a bovine phenol UDP-glucuronosyltransferase, BovUGT1A6.
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cDNA cloning and expression of a bovine phenol UDP-glucuronosyltransferase, BovUGT1A6.

机译：牛酚UDP-葡糖醛酸糖基转移酶BovUGT1A6的cDNA克隆和表达。

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摘要

A full-length cDNA encoding a phenol UDP-glucuronosyltransferase was isolated by plaque hybridization, RT-PCR and 5'-RACE from a cDNA library prepared from the bovine liver. The deduced amino acid sequence (529 amino acid residues) has A signal sequence (23 amino acid residues) at the amino terminus and a transmembrane-anchoring domain (17 amino acid residues) at the carboxyl terminus. The encoded protein has a potential asparagine-linked glycosylation site (Asn291). The cloned cDNA was named bovUGT1A6 on the basis of the amino acid similarity. BovUGT1A6 cloned in the pAAH5 expression vector was transformed into Saccharomyces cerevisiea AH22 cells to obtain an active 54-kDa bovUGT1A6 enzyme. The expressed enzyme represented UDP-glucuronosyltransferase activities toward 1-naphthol and 4-methylumbelliferone, confirming that the isolated cDNA is an isoform of bovine phenol UDP-glucuronosyltransferase. Microsomal UDP-glucuronosyltransferase activity toward 1-naphthol in the bovine kidney cortex was found to be higher than that in the liver and other organs, and mRNA of bovUGT1A6 was more strongly detected in the kidney on Northern blotting analysis. These results suggest that the bovine kidney, which strongly expresses bovUGT1A6, is a significant organ for xenobiotics glucuronidation.
机译：通过噬菌斑杂交，RT-PCR和5'-RACE从牛肝制备的cDNA文库中分离出编码苯酚UDP-葡萄糖醛酸转移酶的全长cDNA。推导的氨基酸序列（529个氨基酸残基）在氨基末端具有A信号序列（23个氨基酸残基），在羧基末端具有跨膜锚定结构域（17个氨基酸残基）。编码的蛋白具有潜在的天冬酰胺连接的糖基化位点（Asn291）。基于氨基酸相似性，克隆的cDNA被命名为bovUGT1A6。将克隆在pAAH5表达载体中的BovUGT1A6转化到酿酒酵母AH22细胞中，获得活性54 kDa bovUGT1A6酶。表达的酶代表UDP-葡萄糖醛糖基转移酶对1-萘酚和4-甲基伞形酮的活性，证实分离的cDNA是牛酚UDP-葡萄糖醛糖基转移酶的同工型。发现牛肾皮质中对1-萘酚的微粒体UDP-葡萄糖醛酸转移酶活性高于肝脏和其他器官中的RNA，并且通过Northern印迹分析在肾脏中更强烈地检测到bovUGT1A6的mRNA。这些结果表明，强烈表达bovUGT1A6的牛肾脏是异生物素葡萄糖醛酸化的重要器官。

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