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。Western blot分析表明。
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.
出处
《淡水渔业》
CSCD
北大核心
2007年第6期65-68,共4页
Freshwater Fisheries
基金
农业部结构调整重大技术研究专项(06-05-03B)资助
