pcDNA3.1-GFP-LC3B真核表达载体的构建  

The Construction of pcDNA3. 1-GFP-LC3B

作　　者：刘宣宣[1,2] 王文倩[1,2] 毛伟平[2] 严浩[2] 孟素芳 王芳[2] 

机构地区：[1]南京医科大学康达学院生理学教研室,江苏连云港222000 [2]南京师范大学江苏省分子医学实验室,江苏南京210023

出　　处：《安徽农业科学》2015年第6期82-84,97,共4页Journal of Anhui Agricultural Sciences

摘　　要：[目的]构建含GFP-LC3B基因的真核表达载体。[方法]从HEK293细胞中提取总RNA,以反转录得到c DNA为模板,根据Gen Bank公布的人源LC3B基因序列设计引物,通过PCR方式扩增目的基因,将基因和pc DNA3.1-GFP载体双酶切后,通过T4连接酶将基因连接到pc DNA3.1-GFP载体上,得到pc DNA3.1-GFP-LC3B真核表达载体。[结果]扩增出目的基因,构建了含目的基因的真核表达载体pc DNA3.1-GFP-LC3B。[结论]成功构建了含有人LC3B基因的真核表达载体pc DNA3.1-GFP-LC3B,为研究细胞自噬提供了基础。[Objective] To construct an eukaryotic expression plasmid expressing GFP-LC3 B gene. [Method]Total RNA was obtained by using Trizol reagent extraction from HEK293 cells. Then c DNA was obtained by reverse transcription,using which as template for PCR amplification of LC3 B gene fragment. The LC3 B gene fragment was inserted into the eukaryotic expression vector pc DNA3. 1-GFP. [Result]LC3B gene was correctly amplified. The recombinant eukaryotic expression plasmid pc DNA3. 1-GFP-LC3 B was successfully constructed. [Conclusion] This recombinant plasmid pc DNA3. 1-GFP-LC3 B will provide a basis for further study on autophagy.

关 键 词：HEK293 CELLS LC3B pcDNA3.1-GFP 真核表达 

分 类 号：S188[农业科学—农业基础科学]