箭筈豌豆甘油醛-3-磷酸脱氢酶基因片段的克隆及序列分析

Cloning and sequencing of GAPDH gene from Vicia sativa

本研究预期通过同源克隆获得了箭筈豌豆(Vicia sativa)甘油醛-3-磷酸脱氢酶(GAPDH)基因的部分片段,为研究箭筈豌豆适应高海拔环境的分子机制奠定基础。根据近缘物种GAPDH基因序列设计1对引物,通过RT-PCR技术克隆获得了一段长度1 141 bp的序列。生物信息学分析表明,该序列编码的381个氨基酸,与其他植物同源GAPDH基因序列的相似度达83%,氨基酸序列相似度在95%以上。可以确定,克隆获得的片段即为箭筈豌豆甘油醛-3-磷酸脱氢酶基因片段。将此片段命名为VsGAPDH,在GenBank注册,登录号HM117006。

To provide molecular basis of high-altitude adaption, a partial sequence of glyceraldehyde-3- phosphate dehydrogenase （GAPDH） gene was isolated from common vetch （Vicia sativa）. In this study, a pair of primers was designed according to homologous GAPDH genes among closely related plants, and then the reverse transcription PCR （RT-PCR） was performed. A nucleotide sequence of 1 141 bp with coding 381 animo acides was identified. Homological analysis showed that the similarity between this sequence and GAPDH gene sequences of other organisms was over 83% in nucleoride sequence level and over 95% in animo acid level. Therefore, the sequence was considered to be V. sativa GAPDH gene. It was named VsGAPDH, and was submited to GenBank with accession number HMl17006.