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本研究基于电子克隆获得的大豆GmASADH基因cDNA序列,根据其编码区设计特异引物,以大豆品种小鹦哥豆总RNA为模板,通过RT-PCR获得了约1 130bp的cDNA片段,T/A克隆后进行序列测定。测序结果显示,GmASADH基因家族有两个成员,命名为GmASADH1(FJ581425)和GmASADH2(HM015631)。GmASADH1编码区长度为1 134bp,编码378个氨基酸,由7个外显子和6个内含子构成,定位于Gm08染色体。GmASADH2编码区长度为1 131bp,编码377个氨基酸,由7个外显子和6个内含子构成,定位于Gm05染色体。两个基因在氨基酸水平上的相似性达到了96.02%,在二级及三级结构上均有不同程度的差异。ASADH蛋白含有NADB_Rossmann super-family和Semialdhyde_dhC superfamily结构域。
Based on the cDNA sequence of soybean GmASADH obtained by electron cloning, specific primers were designed according to the coding region of the cDNA. Approximately 130 bp cDNA fragment was obtained by RT-PCR using the total RNA of Siberian Budweed as template. The T / A clones After the sequence determination. The sequencing results showed that the GmASADH gene family has two members, named GmASADH1 (FJ581425) and GmASADH2 (HM015631). The coding region of GmASADH1 is 1 134 bp in length and encodes a protein of 378 amino acids. It consists of seven exons and six introns and is located on the Gm08 chromosome. The coding region of GmASADH2 is 1 131bp in length and encodes 377 amino acids. It consists of 7 exons and 6 introns and locates on chromosome Gm05. The similarity of the two genes at the amino acid level reached 96.02%, with varying degrees of secondary and tertiary structure. The ASADH protein contains the NADB_Rossmann super-family and Semialdhyde_dhC superfamily domains.