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[甘草3-羟基-3-甲基戊二酰辅酶A还原酶cDNA的克隆与鉴定]

[Cloning and characterization of 3-hydroxy-3-methylglutary CoA reductase cDNA of Glycyrrhiza uralensis].

作者信息

Rong Qixians, Xu Qiaoxian, Liu Chunsheng, Huang Luqi

机构信息

School of Chinese Materia Medica, Beijing University of Traditional Chinese Medicine, Beijing 100102, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2011 May;36(10):1275-9.

Abstract

OBJECTIVE

To clone and analysis the sequence of 3-hydroxy-3-methylglutary CoA reductase (HMGR) cDNA from Glycyrrhiza uralensis.

METHOD

The primers were designed based on the conservative region of HMGR nucleic acids sequence from public database. The target gene was obtained from root of G. uralensis by use of homologous cDNA amplificati on and RACE technologies. The sequence alignment was performed using BLAST. The open reading frame was identified by use of the ORF Finder. The protein domains were defined by use of Prosite software. Clustal was used to conduct multiple amino acid sequence alignment and MEGA 5.0 was used to conduct the phylogenetic tree.

RESULT

The GuHMGR cDNA sequence was obtained contains 1 842 bp contains a 1 722 bp ORF, encoding 573 amino acids with 3-hydroxy-3-methylglutary CoA reductases family profile. Deduced amino acid sequence had 84% and 76% homology to the amino acid sequence of Pisum sativum, Medicago truncatula.

CONCLUSION

The cloning of 3-hydroxy-3-methylglutary CoA reductase (HMGR) cDNA will provide a foundation for exploring the function of HMGR in glycyrrhizin biosynthesis.

摘要

目的

克隆并分析乌拉尔甘草3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)cDNA序列。

方法

根据公共数据库中HMGR核酸序列的保守区设计引物。利用同源cDNA扩增和RACE技术从乌拉尔甘草根中获得目标基因。使用BLAST进行序列比对。利用ORF Finder鉴定开放阅读框。使用Prosite软件确定蛋白质结构域。使用Clustal进行多个氨基酸序列比对,使用MEGA 5.0构建系统发育树。

结果

获得的乌拉尔甘草HMGR cDNA序列全长1 842 bp,包含一个1 722 bp的开放阅读框,编码573个氨基酸,具有3-羟基-3-甲基戊二酰辅酶A还原酶家族特征。推导的氨基酸序列与豌豆、蒺藜苜蓿的氨基酸序列同源性分别为84%和76%。

结论

3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)cDNA的克隆为探索HMGR在甘草酸生物合成中的功能奠定了基础。

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