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利用噬菌体随机十二肽库筛选生长抑素抗原模拟表位

Screening the Mimotopes of Porcine Somatostatin with Phage Display 12-mer Random Peptide Library

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【Author】 YUAN Fang-yan~(1))HE Yu-long~(1))ZHAO Na~(1))NIE Fen~(1)) WU Xiao-xiong~(2))LONG Liang-qi~(2))LI Kui~(3))LUO Xiao-song~(4))(~(1)) College of Fisheries,Huazhong Agricultural University,Wuhan 430070,China;~(2))College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070,China;~(3))Institute of Animal Science,Chinese Academy of Agriculture Sciences,Beijing 100094,China;~(4)) Yangtze River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Jingzhou 434000,China)

【机构】华中农业大学水产学院；华中农业大学动物医学院；中国农业科学院畜牧研究所；中国水产科学研究院长江水产研究所武汉430070；武汉430070；北京100094；荆州434000；

【摘要】用生长抑素(somatostatin,SS)疫苗“激生1号”免疫日本大耳兔得到抗血清,经饱和硫酸铵粗提IgG,再经DEAE-sephadex-A50低压层析系统纯化后,以此IgG作为筛选配基,对随机十二肽库进行4轮淘洗。随机挑取22个噬菌斑进行双夹心ELISA,结果有6个克隆与SS抗体有较强的特异性结合力,竞争抑制ELISA进一步证明6个克隆有较好的抑制作用。将6个阳性克隆提取ssDNA进行测序,测序结果中有5个克隆的外源肽序列完全一致,经与SS氨基酸序列比对,发现两者有共同序列FWK,说明5个克隆模拟了生长抑素的抗原表位。更多还原

【Abstract】 The sera were obtained by immunizing the rabbit with somatostatin vaccine.The IgG rudely purified by saturated ammonia sulphate was purified through DEAE-sephadex-A50 low pressure chromatography system.Using the IgG as target,a 12-mer random peptide library was biopanned for 4 rounds.The 22 clones eluted from 4rd panning were picked out randomly and characterized.Six positive clones among 22 clones were identifited by sandwith ELISA and competition ELISA.The ssDNA of the 6 positive clones were abstracted and sequenced.The results showed that five clones had the same sequence.Compared with SS amino acid sequence,the FWK was their homogerous.更多还原

【关键词】生长抑素；噬菌体随机肽库；模拟表位；
【Key words】 somatostatin； phage display random peptide library； mimotope；

【基金】国家“863”项目(2001AA222241)资助

【文献出处】华中农业大学学报 ,Journal of Huazhong Agricultural University(Natural Science Edition) , 编辑部邮箱 ,2006年06期

【分类号】S852.5
【下载频次】128

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