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猪囊尾蚴cDNA 质粒表达文库的构建

宗瑞谦1，2， 才学鹏1， 季建莉3， 景志忠1

（1. 中国农业科学院 兰州兽医研究所 农业部畜禽病毒学重点开放实验室， 甘肃 兰州 730046； 2. 甘肃农业大学 动物医学院， 甘肃 兰州 730070； 3. 新疆农业职业技术学院 动物科学系， 新疆 昌吉 831100）

摘要： 从猪囊尾蚴中提取总RNA，并分离出mRNA，采用定向克隆方法，将猪囊尾蚴cDNA片段重组入质粒表达载体pSPORT1的 NotⅠ和Sal Ⅰ双酶切位点之间，构建了猪囊尾蚴cDNA表达文库。通过库容量鉴定，所构建的表达文库的容量为2×106；经含有IPTG和Xgal的颜色选择平皿测定，其重组率为100％。

关键词： 猪囊尾蚴； cDNA表达文库； 构建

中图分类号：S 852.734  文献标识码： A  文章编号：1000-6419(2004)05-0011-03

Construction of cDNA library from Cysticercus cellulosae

ZONG Ruiqian1,2， CAI Xuepeng1, JI Jianli3, JING Zhizhong1
(1. Key Laboratory of Animal Virology, Ministry of Agriculture/Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046,China;
 2. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070,China; 
3. Department ofAnimal Science, Xinjiang Agricultural Vocational Technological College, Changji 831100,China)

Abstract: The total RNA was extracted and mRNA was isolated from Cysticercus cellulosae . The doublestranded cDNA was synthesized and cloned into pSPORTⅠ vector using SuperscriptTMPlasmidSystem with GATEWAYTM Technology for cDNA Synthesis and Cloning kit offered by Invitrogen Corporation. The results showed that the recombinant rate was 100％ and the library contained 2×106recombinants. It was proved that the cDNA expressing library was constructed successfully with the directional cloning method.

Key words: Cysticercus cellulosae ; cDNA expressing library; construction

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