木薯MinE2基因的亚细胞定位和表达分析

MeMinE2蛋白是一种亲水性蛋白,在Min系统调控质体分裂环过程中,MinE2蛋白能够与其他蛋白相互作用协调FtsZ环的定位。为验证木薯MinE2基因的功能,从木薯基因组中分离出全长为696 bp的MinE2基因。用Protparam在线软件对Me MinE2蛋白做相关分析,该蛋白由231个氨基酸组成,其GRAVY值为-0.256,是一个不稳定的亲水性蛋白。构建1300-GFP-MinE2瞬时表达载体,然后转染原生质体,用激光共聚焦扫描显微镜观察观察原生质体的形态,亚细胞定位显示MinE2蛋白定位于叶绿体中。构建pET28aMinE2原核表达载体,并转化至E. coli BL21 (DE3),经8 mmol/L IPTG诱导MeMinE2蛋白过表达,大肠杆菌异常分裂。本研究对MeMinE2蛋白功能的解析,为进一步探索MinE2基因对木薯质体分裂和植物育种的影响提供一定的理论依据。

Subcellular Localization and Expression Analysis of MinE2 Gene in Cassava

MeMine2 protein is a kind of hydrophilic protein.MeMine2 protein can interact with other proteins to coordinate the location of FtsZ ring in the process of Min system regulating plastid division ring.In order to verify the function of MinE2 gene in cassava,a total length of 696 bp MinE2 gene was isolated from the cassava genome.The online software proteparam showed that the protein is composed of 231 amino acids,and it is an unstable hydrophilic protein with gray value of-0.256.Construction of 1300-GFP-Mine2 transient expression vector and transfection of protoplast.Observation of protoplast morphology with laser confocal scanning microscope.Subcellular localization showed that Mine2 protein is located in chloroplast.Construction of PET28 a-Mine2 prokaryotic expression vector and transformation to E.coli BL21(DE3).Abnormal division of E.coli in overexpression of MeMine2 protein induced by 8 mm IPTG.This study provides a theoretical basis for further study on the effect of MinE2 gene on the plastid division and plant breeding of cassava.