花生羧酸酯酶AhCXE的克隆与原核表达

Clone and Prokaryotic Expression of Carboxylesterase AhCXE in Peanut

前期功能研究发现当倒置离体培养的果针发生向地性弯曲时,花生羧酸酯酶(AhCXE)基因表达量显著增加。本研究以花生果针为材料,利用RT-PCR与RACE克隆技术,克隆了AhCXE基因并通过生物信息学技术分析了AhCXE基因的基本特征,利用原核诱导技术分析目标蛋白的结构与蛋白表达特征。测序结果表明AhCXE基因ORF全长为1 068 bp,编码355个氨基酸;生物信息学分析显示AhCXE蛋白属于α/β蛋白水解酶家族,具有酯酶活性,参与体内脂类代谢,与大豆羧酸酯酶具有较高的相似度,最后将该蛋白编码的基因命名为AhCXE。

Previous studies have found that when peanut peg cultured in vitro was bended geotropically, the expression quantity of peanut carboxylesterase （A hCXE） gene increased significantly. This study used peanut peg as materials, cloned gene A hCXE by RT-PCR and PACE clone technology, analyzed its essential characteristic through bioinformatics technology, and analyzed the strucatre and protein expression of the target protein by original induced nuclear technology. Sequencing results showed that the ORF of AhCXE was 1 068 bp in full length, encoding 355 amino acids. Bioinformatics analysis indicated that the protein AhCXE belonged to α/β proteolytic enzymes family, which possessed esterase activity, involved in lipid metabolism in the body, and had high similarity with soybean carboxylesterase, based on which we named this protein coding gene as A hCXE.