LLC-PK1细胞中LDHB蛋白的体外表达及小RNA干扰  

In vitro expression of LDHB in LLC-PK1 cells and its sRNA interfering

作　　者：罗依然[1] 周书亭 杨亮 刘源平 蒋升瑶 叶力波利·达吾力 王子泓 罗渝宵 邹霭萱 崔立[1] 杨志彪[1] LUO Yi-ran;ZHOU Shu-ting;YANG Liang;LIU Yuan-ping;JIANG Sheng-yao;YELIBOLI Dawuli;WANG Zi-hong;LUO Yu-xiao;ZOU Ai-xuan;CUI Li;YANG Zhi-biao(Shanghai Key Laboratory of Veterinary Biotechnology/School of Agriculture and Biology,Shanghai Jiaotong University,Shanghai 200240,China)

机构地区：[1]上海交通大学农业与生物学院/上海市兽医生物技术重点实验室,上海200240

出　　处：《上海交通大学学报（农业科学版）》2019年第6期87-93,共7页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：国家重点研发计划(2016YFD0500100);上海市科技兴农项目[沪农科推字(2017)第1-11号];国家自然科学基金(31472211)。

摘　　要：为了观察与猪流行性腹泻病毒(PEDV)NSP12互作蛋白乳酸脱氢酶B(LDHB)的体外表达情况和小RNA干扰效果,构建了真核表达载体pcDNA4.0-LDHB-3*Flag,并在Vero细胞中过表达LDHB,发现在37 kDa处有大小相符的明显条带。设计的2条小干扰RNA能降低LLC-PK1细胞中LDHB的mRNA和蛋白质水平,经one-way ANOVA检验证明干扰实验组2-ΔΔct的平均值显著低于对照组(P<0.05)。该结果为研究LDHB在PEDV感染LLC-PK1和Vero细胞过程中的作用提供了基础数据。In order to observe the in-vitro expression of LDHB interacting with PEDV NSP12 and the efficiency of the small interfering RNA,a eukaryotic expression vector named pcDNA4.0-LDHB-3*Flag was constructed to over-express LDHB in Vero cells,and a matching protein band was detected at 37kDa.The mRNA and protein levels of LDHB were significantly down-regulated in LLC-PK1 cells after interfered by two small RNAs.One-way ANOVA test showed the mean of 2-ΔΔct of experiment groups was substantially below the control groups(P<0.05).This result provided a basis for the functional analysis of LDHB in the process of PEDV infecting LLC-PK1 and Vero cells.

关 键 词：LDHB 真核表达 小RNA干扰 

分 类 号：S828[农业科学—畜牧学]