表达驱动蛋白KIF4A磷酸化突变体蛋白重组杆状病毒的构建  

Construction of Recombinant Baculovirus Expressing KIF4A Phosphorylation Mutant Proteins

作　　者：魏雅婷 李心怡 朱长军[1,2] WEI Yating;LI Xinyi;ZHU Changjun(College of Life Sciences,Tianjin Normal University,Tianjin 300387,China;Tianjin Key Laboratory of Animal and Plant Resistance,Tianjin 300387,China)

机构地区：[1]天津师范大学生命科学学院,天津300387 [2]天津动植物抗性重点实验室,天津300387

出　　处：《生物化工》2024年第4期49-52,57,共5页Biological Chemical Engineering

基　　金：天津市多元投入基金重点项目(21JCZDJC00960)。

摘　　要：目的:探究驱动蛋白家族成员4A(KIF4A)在细胞有丝分裂早期调控染色质凝缩的作用机制。方法:本研究构建表达KIF4A磷酸化突变体蛋白的重组质粒,转化DH10Bac感受态细胞,蓝白斑筛选后提取重组杆状病毒基因组,转染Sf9细胞并培养以获得病毒液。结果:通过昆虫杆状病毒表达系统成功获得表达KIF4A突变体蛋白的病毒液。结论:本研究为后续大量表达及体外纯化KIF4A突变体蛋白奠定了实验基础。Objective:Exploring the mechanism of kinesin family member 4A(KIF4A) in regulating chromatin condensation during early cell mitosis.Methods:In this study,a recombinant plasmid expressing KIF4A phosphorylated mutant protein is constructed and transformed into DH10Bac competent cells.After blue and white spot screening,the recombinant baculovirus genome is extracted,transfected into Sf9 cells,and cultured to obtain virus solution.Results:The viral solution expressing KIF4A mutant protein is successfully obtained through the insect baculovirus expression system.Conclusion:This study lays the experimental foundation for the subsequent large-scale expression and in vitro purification of KIF4A mutant protein.

关 键 词：驱动蛋白 KIF4A 点突变 杆状病毒 

分 类 号：Q789[生物学—分子生物学]