木薯RZFP锌指蛋白的基因克隆及其蛋白原核表达
Gene Cloning and Protein Prokaryotic Expression of RZFP Zinc Finger Protein in Cassava
葛中元 1赵惠萍1
作者信息
- 1. 海南大学热带作物学院,海南省耐盐作物生物技术重点实验室,海口,570228;海南大学热带作物学院,南繁与热带高效农业协同创新中心,海南省热带生物资源可持续利用重点实验室-省部共建国家重点实验室培育基地,海口,570228
- 折叠
摘要
锌指蛋白在植物生长发育以及适应环境胁迫中发挥了重要的作用,为了更好地研究锌指蛋白在木薯的生长发育以及抗逆境胁迫中的功能.本研究通过分离鉴定并克隆了一个MeRZFP基因,测序结果显示该基因大小为1068bp.首先对其进行了系统进化树和保守结构域分析,发现其含有zinc-ribbon和zinc finger-C3C4这两个保守结构域.进一步构建了原核表达载体pET28a-MeRZFP,筛选出了相对适合其菌株生长的温度,结果表明该蛋白最适表达温度为28 ℃,并且对重组蛋白pET28a-MeRZFP进行SDS-PAGE电泳检测以及Western-blot验证,结果显示该蛋白大小为39 kD.为进一步探究木薯RZFP在木薯应对环境胁迫时发挥的功能提供了一定的参考.
Abstract
Zinc finger proteins play an important role in plant growth and development and adaptation to enviro-nmental stress.In order to better study the function of zinc finger protein in cassava growth and development and resistance to stress.In this study,a MeRZFP gene was isolated,identified and cloned.The sequencing results showed that the gene size was 1 068 bp.Firstly,the study analyzed its phylogenetic tree and conservative domain,and found that it contains two conservative domains:zinc-ribbon and zinc-finger-C3C4.The prokaryotic expression vector pET28a-MeRZFP was further constructed,and the temperature that was relatively suitable for the growth of the strain was screened.The results showed that the optimal expression temperature of the protein was 28 ℃.The recombinant protein pET28a-MeRZFP was detected by SDS-PAGE electrophoresis and verified by Western blot.The results showed that the size of the protein was 39 kD.It provides a reference for further exploring the function of cassava RZFP in cassava's response to environmental stress.
关键词
木薯/RING型锌指蛋白/基因克隆/原核表达Key words
Cassava(Manihot esculenta)/RING zinc-finger protein/Gene cloning/Prokaryotic expression引用本文复制引用
基金项目
海南省研究生创新科研课题(Qhyb2021-27)
海南大学教育教学改革研究项目(HDJG-Y202220)
出版年
2024
NETL
NSTL
维普
万方数据