《植物生理学报》 2018, 54(3): 441-448

一个麻疯树花羧酸酯酶基因的克隆、表达分析和原核表达

雷世康^1,2, 徐刚^1,3,*

¹贵州省森林资源与环境研究中心, 贵阳550025; ²贵州大学生命科学学院, 贵阳550025; ³贵州大学昆虫研究所, 贵阳550025

通信作者：徐刚；E-mail: xg335300@aliyun.com

摘　要：

基于麻疯树RNA-Seq数据, 利用RT-PCR技术成功从麻疯树花器官中克隆获得JcCXE7基因的开放阅读框, 长度为957 bp, 编码318个氨基酸, 预测该蛋白分子质量为35.55 kDa。对JcCXE7基因的时空表达进行分析, 结果表明, JcCXE7基因可能参与了麻疯树雌花大孢子母细胞发生和减数分裂; 且在雌花中的表达水平大大高于雄花。利用原核表达体系对JcCXE7基因进行诱导表达, 随后对获得的蛋白进行了LC-MS/MS鉴定, 并对Jc-CXE7蛋白的氨基酸序列与同源家族进行多重比对及其结构进行分析, 结果表明JcCXE7蛋白可能为GID1家族蛋白。

关键词：麻疯树; 花发育; 羧酸酯酶; 表达分析

收稿：2017-08-14   修定：2018-01-18

资助：国家自然科学基金(31460184)。

Gene cloning, expression pattern analysis and prokaryotic expression of a carboxylesterase in flower of Jatropha curcas

LEI Shi-Kang^1,2, XU Gang^1,3,*

¹Institute for Forest Resources and Environment of Guizhou, Guizhou University, Guiyang 550025, China; ²College of Life Sciences, Guizhou University, Guiyang 550025, China; ³Institute of Entomology, Guizhou University, Guiyang 550025, China

Corresponding author: XU Gang; E-mail: xg335300@aliyun.com

Abstract:

A sex differently expressed carboxylesterase gene named as carboxylesterase 7 (JcCXE7) was screened out from transcriptome of flower buds in Jatropha curcas. The open reading frame (ORF) of JcCXE7 was obtained using RT-PCR. Sequencing result showed that the ORF of JcCXE7 was 957 bp long, and predicted to encode a piptide of 318 amino acids with a molecular mass of 35.55 kDa. Expression pattern analysis of JcCXE7 gene during flower development was analysis using qRT-PCR. After sex differentiation, JcCXE7 was firstly up-regulated in the stage of macrospore mother cell in female, followed by a gradually down-regulation; while it was greatly down-regulated in the stage of microspore mother cell in male with a moderate up-regulation at the stage of pollen maturation. Overall, the expression level of JcCXE7 was higher in female than that in male during the flower development. The prokaryotic expression production of JcCXE7 was identified and analyzed by LC-MS/MS. The multiple sequence alignment and supersecondary structure analysis of JcCXE7 protein suggested that it would be GID1 family protein.

Key words: Jatropha curcas; flower development; carboxylesterase; expression analysis