[1]张健,陈艳,乔传玲,等.表达猪白细胞介素2重组腺病毒的构建[J].中国预防兽医学报,2011,(04):313-315.
 ZHANG Jian,CHEN Yan,QIAO Chuan-ling*,et al.Construction of recombinant adenovirus expressing porcine interleukin 2 gene[J].Chinese journal of preventive veterinary medicine,2011,(04):313-315.
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表达猪白细胞介素2重组腺病毒的构建
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2011年04期
页码:
313-315
栏目:
研究简报
出版日期:
2011-04-15

文章信息/Info

Title:
Construction of recombinant adenovirus expressing porcine interleukin 2 gene
作者:
张健;陈艳;乔传玲;杨焕良;唐续;辛晓光;陈化兰;
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;
Author(s):
ZHANG Jian CHEN Yan QIAO Chuan-ling* YANG Huan-liang TANG Xu XIN Xiao-guang CHEN Hua-lan
Animal Influenza Laboratory of the Ministry of Agricultural, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China
关键词:
IL-2重组腺病毒基因表达
Keywords:
interleukin 2 recombinant adenovirus gene expression
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。
Abstract:
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

参考文献/References:

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备注/Memo

备注/Memo:
基金:哈尔滨市科技攻关计划(2009AA6BN078)
更新日期/Last Update: 2011-09-29