表达猪白细胞介素2重组腺病毒的构建  

Construction of recombinant adenovirus expressing porcine interleukin 2 gene

作　　者：张健[1] 陈艳[1] 乔传玲[1] 杨焕良[1] 唐续[1] 辛晓光[1] 陈化兰[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/农业部动物流感重点开放实验室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2011年第4期313-315,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金：哈尔滨市科技攻关计划(2009AA6BN078)

摘　　要：为构建表达猪白细胞介素2(IL-2)的复制缺陷型重组腺病毒,本研究提取经刀豆球蛋白A(Con A)刺激体外培养的长白猪外周血淋巴细胞总RNA,利用RT-PCR扩增猪IL-2基因,并克隆于质粒pIRES2-EGFP中,将含有IL-2及EGFP的基因片段亚克隆到腺病毒的穿梭质粒pDC315中,构建重组腺病毒穿梭质粒pDC315-IL-2-EGFP。利用脂质体转染方法将pDC315-IL-2-EGFP和腺病毒骨架质粒pBHGloxΔE1,E3Cre共转染HEK293细胞。根据腺病毒感染后形成的典型细胞病变及EGFP的荧光信号筛选重组腺病毒rAd-IL-2-EGFP。经western blot鉴定,结果表明获得了一株能够表达猪IL-2蛋白的重组腺病毒rAd-IL-2-EGFP。本研究为进一步研究猪IL-2在疫苗免疫中的增强作用及研制新型免疫增强剂奠定了基础。To construct a recombinant adenovirus expressing porcine interleukin 2 （IL-2）, The IL-2 gene was amplified by RT-PCR with specific primers from the total RNA, which were extracted from peripheral blood lymphocytes after stimulation with ConA in vitro and cloned into pIRES2-EGFP. The recombinant adenovirus shuttle plasmid pDC315-IL2-EGFP was constructed by inserting IL2-IRES-EGFP into shuttle plasmid pDC315. A recombinant adenovirus expressing IL-2 （rAd-IL-2-EGFP） was generated by co-transfecting the pDC315-IL2-EGFP and the backbone plasmid pBHGlox△E1, E3Cre in HEK293 cells. The rAd-IL2-EGFP exhibited the typical cytopathic effect of adenovirus and fluorescence expressed by EGFP gene in HEK293 cells and IL-2 expression was confirmed by western blot. Therefore, rAd-IL-2-EGFP would be useful to study the immune enhancement of the porcine IL-2 in pig.

关 键 词：IL-2 重组腺病毒 基因表达 

分 类 号：S852.65[农业科学—基础兽医学]