水稻中编码甘油-3-磷酸转酰酶部分cDNA的克隆及序列分析

Cloning and Sequencing of a Part-Length cDNA Coding forGlycerol-3-Phosphate Acyltransferase from Rice

参照国外报道的几种双子叶植物的甘油－３－磷酸转酰酶的相对保守的氨基酸序列，设计并合成一对简并引物，提取抗冷性强的水稻品种“丽梗２号”的ＲＮＡ，采用ＲＴ－ＰＣＲ技术，扩增出了３１５ｂｐ的一段ｃＤＮＡ片段。扩增产物经纯化后直接克隆到ｐＧＥＭ－Ｔ载体系统中，经ＰＣＲ法鉴定，所得的重组质粒中含有３１５ｂｐ的片段。证明已克隆到水稻甘油－３－磷酸转酰酶部分ｃＤＮＡ。采用双链双脱氧法定序分析，表明所克隆的该段ｃＤＮＡ序列与国外所报道的双子叶植物的该段ｃＤＮＡ的核苷酸序列及氨基酸序列的同源性均达到７０％以上，提示了该基因在进化上具有一定的保守性。

According to the conserved amino acid sequences of glycerol-3-phosphate acyltransferase from several kinds of dicotyls, we designed and synthesized a pair of degenerate primers. RNA was isolated from the highly chilling-resistant rice “Ligeng No.2”. A 315bp cDNA fragment was obtained with RT-PCR technique. After purification, the cDNA fragment was cloned into pGEM-T vector directly. The result of identification by PCR method indicates that the recombinant plasmid pGTL5 contains a 315bp cDNA fragment. The result demonstrated that we had cloned a part of cDNA encoding glycerol-3-phosphate acyltransferase. While the cDNA sequence and deduced amino acid sequence reported here was compared with that published abroad, the identity was above 70%. This result hints that the gene is highly conservative on evolution.