华支睾吸虫cDNA文库的构建  

Construction of cDNA Library from Clonorchis sinensis

作　　者：吴军[1] 裴福全[1] 钟肖芬[2] 卫剑文[2] 彭立胜[2] 阮彩文[1] 崔惠儿[1] 方悦怡[1] 潘波[1] 

机构地区：[1]广东省寄生虫病防治研究所,广州510300 [2]中山大学生命科学学院

出　　处：《中国人兽共患病杂志》2003年第4期91-92,共2页Chinese Journal of Zoonoses

基　　金：广东省卫生厅科研经费资助 (A19990 81)

摘　　要：目的　构建华支睾吸虫cDNA文库。方法　采集华支睾吸虫阳性鱼 ,分离囊蚴 ,感染实验动物兔 ,解剖兔收集华支睾吸虫成虫虫体。应用“一步法”提取华支睾吸虫总RNA ;经过mRNA纯化、cDNA合成 ,以PcDNA3(Amp +)质粒为载体构建文库。挑取 2 0个克隆进行扩增 ,选择 9个克隆进行DNA序列测定。序列结果与GenBank中相关基因进行比对。结果　获得含有 9 7× 10 5个重组子库容量的华支睾吸虫cDNA文库。其中PC6号克隆基因序列与华支睾吸虫半胱氨酸蛋白酶序列具有 93%的同源性。结论　已构建成华支睾吸虫cDNA文库。Aim To constructe the cDNA library of C sinenis Methods Collected the bladder larva from fish,infected the rabbits Gained the imagines of C sinenesis from the livers of the rabbits after three months Using the 'One step method' extracted the total RNA Purified mRNA and synthesized cDNA and constructed the cDNA library with the PcDNA3(Amp+) Picked out 20 clones to multiply 9 clones had been sequenced and compared the similarity with 'GENBANK' Results Obtained the cDNA library of C sinensis that contained 9 7×10 5 recombine clones The PC6 clone has 93% similarity with the cysteine enzyme of C sinensis Conclusion cDNA library for C sinensis has been constructed.

关 键 词：华支睾吸虫 CDNA文库 构建 克隆基因序列 

分 类 号：R383.22[医药卫生—医学寄生虫学]