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广西产眼镜王蛇毒α-神经毒素基因的分子克隆及序列分析
引用本文:李其斌,黄光武,赵晓琴,渡庆次贺博,金城记代彦,中村真理子,小杉忠诚.广西产眼镜王蛇毒α-神经毒素基因的分子克隆及序列分析[J].广西科学,2002,9(3):202-206.
作者姓名:李其斌  黄光武  赵晓琴  渡庆次贺博  金城记代彦  中村真理子  小杉忠诚
作者单位:1. 广西医科大学第一附属医院急诊科蛇伤救治与蛇毒临床应用研究中心,南宁市滨湖路6号,530021
2. 日本琉球大学医学部生理学科
摘    要:为获得眼镜王蛇(Ophiophagus hannah,简称Oh)蛇毒α-神经毒素(α-NT)的基因序列,依据眼镜蛇科不同毒蛇种类来源的α-NT基因有较高的同源性,设计1对上下游引物,为克服引物带来模糊扩增,在蛋白编码部分再设计1对上下游特异引物,用Nacleospin RNA Kit法从3条活眼镜王蛇毒腺中提取mRNA,以3′端引物合成的cDNA作为模板进行PCR扩增反应,测定产物的核苷酸序列,得到全长474bp的眼镜王蛇cDNA基因核苷酸序列。该核苷酸序列的信号肽与眼镜蛇树属Pseudonnaja textilis(Pt)、海蛇Laticauda semifasciata(Ls)100%同源,与眼镜蛇南洋亚种Naja sputatrix (Ns)、银环蛇(Bungarus multicinctus)(Bm)96.8%同源;蛋白密码部分有83.3%与Ns、79.2%与Pt、76.4%与Ls、74.1%与Bm同源。信号肽后紧接着的72个氨基酸有90.3%与已发现的眼镜王蛇毒长链α-NT Toxin a同源,大约有73.6%与Toxin b、69.7%与Oh-4、66.7%与Oh-5、56.9%与Oh-6A和6B同源,并与α-银环蛇毒素54.2%同源。说明新发现的眼镜王蛇cDNA是一条长链α-NT基因。

关 键 词:蛇毒α-神经毒素  分子克隆  眼镜王蛇  基因序列
收稿时间:2001/2/1 0:00:00
修稿时间:2001年2月1日

Cloning and Sequence Analysis of the cDNA Encoding of α-Neurotoxin from Guangxi King Cobra (Ophiophagus hannah)
Li Qibin,Huang Guangwu,Zhao Xiaoqin,Y. Togesi,K. Kinjoh,M. Nakamura and T. Kosugi.Cloning and Sequence Analysis of the cDNA Encoding of α-Neurotoxin from Guangxi King Cobra (Ophiophagus hannah)[J].Guangxi Sciences,2002,9(3):202-206.
Authors:Li Qibin  Huang Guangwu  Zhao Xiaoqin  Y Togesi  K Kinjoh  M Nakamura and T Kosugi
Institution:Snakebite Research Department, The first Affiliated Hospital, Guangxi Medical University, 6 Binhulu, Nanning, Guangxi, 530021, China,Snakebite Research Department, The first Affiliated Hospital, Guangxi Medical University, 6 Binhulu, Nanning, Guangxi, 530021, China,Snakebite Research Department, The first Affiliated Hospital, Guangxi Medical University, 6 Binhulu, Nanning, Guangxi, 530021, China,Snakebite Research Department, The first Affiliated Hospital, Guangxi Medical University, 6 Binhulu, Nanning, Guangxi, 530021, China,Snakebite Research Department, The first Affiliated Hospital, Guangxi Medical University, 6 Binhulu, Nanning, Guangxi, 530021, China,Snakebite Research Department, The first Affiliated Hospital, Guangxi Medical University, 6 Binhulu, Nanning, Guangxi, 530021, China and Snakebite Research Department, The first Affiliated Hospital, Guangxi Medical University, 6 Binhulu, Nanning, Guangxi, 530021, China
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