大豆乙酰羟基酸异构还原酶基因GmAHRI的克隆与分析

Molecular Cloning and Analysis of GmAHRI Gene in Soybean

基于电子克隆获得的大豆GmAHRI基因cDNA序列编码区设计特异引物,以高异亮氨酸大豆品种"和龙早熟豆"总RNA为模板,通过RT-PCR获得了约1 764 bp的cDNA片段,T/A克隆后进行序列测定。测序结果显示:GmAHRI基因家族有2个成员,分别命名为GmAHRI1(FJ594399.1)和GmAHRI2(JN034043)。GmAHRI1和GmAHRI2的编码区长度均为1 764 bp,编码587个氨基酸,由10个外显子和9个内含子构成,分别定位于Gm12和Gm13染色体上。2个基因在氨基酸水平上的相似性达到了96.71%,所编码的蛋白质在二级结构上有所差异,三级结构相同。

In this research,the GmAHRI gene was cloned from Glycine max and analyzed its sequences.A cDNA product about 1764 bp was amplified from the total RNA of soybean leaves by reverse transcription PCR（RT-PCR）with specific primers designed with the full-length sequence of GmAHRI gene.Sequence analysis showed that the GmAHRI gene family had two members,named GmAHRI1（FJ594399.1）and GmAHRI2（JN034043）,respectively.The full length of GmAHRI1 and GmAHRI2 were 1764 bp,encoding 587 amino acids,constructed by 10 exons and 9 introns,and located on chromosome Gm12 and Gm13,respectively.The similarity of two genes reached to 96.71% in amino acid level,there were obvious difference in secondary structure,but they had same tertiary structure.