学术
鲤胰岛素样生长因子Ⅱ的克隆表达与抗体制备
商汉桥[1] 罗晓松[1,2] 邹世平[1,2] 伍晓雄[1] 吴小平[1] 龙良启[1]
[1]华中农业大学,武汉430070 [2]中国水产科学研究院长江水产研究所,湖北荆州434000
摘 要:
从鲤（Cyprinus carpio）肝脏总RNA中成功地克隆和诱导表达了胰岛素样生长因子Ⅱ（IGF-Ⅱ）基因，并用间接ELISA法和Western-blot对其融合蛋白的多克隆抗体的效价和特异性进行了分析。结果表明，克隆的鲤IGF-Ⅱ基因与已报道的序列相比，有1个位点发生同义突变；经SDS-PAGE电泳分析，目的蛋白主要以包涵体的形式存在，表达量占菌体总量的35％。经ELISA测定，制备的抗体效价为6400。Westernblot分析表明，抗血清能与重组蛋白发生特异性反应。[著者文摘]
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文章出处:
《淡水渔业》-2007年37卷6期 -65-68页
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文献标识码:
A
文章编号:
1000-6907-（2007）06-0065-04
Gene Cloning and Fusion Expression of Common Carp Insulin-like Growth Factor Ⅱ and Preparation of Its Polyclonal Antibody
SHANG Han-qiao , LUO Xiao-song, ZOU Shi-ping,WU Xiao-xiong , WU Xiao-ping , LONG Liang-qi（ 1. Huazhong Agricultural University, Wuhan 430070; 2. Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Jingzhou, Hubei 434000）
Abstract:
Common carp insulin-like growth factors （IGF-Ⅱ ）gene was successfully cloned from liver total RNA, and expressed by inducing. The titer and specificity of the rabbit anti-IGF Ⅱ pelyclonal antibody of fusion protein was tested by ELISA and Western blot. The result showed that there was a synonymous mutation of 1 site compared with sequence reported. The SDS-PAGE showed that the recombinant protein expressed in the form of inclusion bodies and amounted to 35% of the whole protein in the E. coli cell. ELISA showed that the titer of the antiserum was 6400. Western blot indicated that the antiserum could react to the recombinant protein specifically.[著者文摘]
Key words:
common carp（ Cyprinus carpio） ; IGF-Ⅱ ; cloning; prokaryotic expression; polyclonal antibody
基金资助:
农业部结构调整重大技术研究专项（06-05-03B）资助
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