王朝兰, 汤冬生, 王业梅, 姚勇, 汪学龙. 猪囊尾蚴GP50编码基因的克隆和序列分析[J]. 蚌埠医学院学报, 2012, 36(3): 256-259.
    引用本文: 王朝兰, 汤冬生, 王业梅, 姚勇, 汪学龙. 猪囊尾蚴GP50编码基因的克隆和序列分析[J]. 蚌埠医学院学报, 2012, 36(3): 256-259.
    WANG Chao-lan, TANG Dong-sheng, WANG Ye-mei, YAO Yong, WANG Xue-long. Cloning and sequencing of GP50 gene from cysticercus cellulosae[J]. Journal of Bengbu Medical College, 2012, 36(3): 256-259.
    Citation: WANG Chao-lan, TANG Dong-sheng, WANG Ye-mei, YAO Yong, WANG Xue-long. Cloning and sequencing of GP50 gene from cysticercus cellulosae[J]. Journal of Bengbu Medical College, 2012, 36(3): 256-259.

    猪囊尾蚴GP50编码基因的克隆和序列分析

    Cloning and sequencing of GP50 gene from cysticercus cellulosae

    • 摘要: 目的:从猪囊尾蚴cDNA中扩增出GP50编码基因,亚克隆至真核表达载体,利用所得的重组蛋白建立一种灵敏检测猪囊尾蚴感染的实验方法。方法:设计并合成引物,从猪囊尾蚴cDNA文库中PCR扩增GP50编码基因,通过与线性克隆载体pGEM-T连接,经酶切和PCR鉴定及DNA测序证实后,软件分析所获的目的基因。结果:PCR法扩增出897 bp左右的DNA片断,将重组质粒pGEM-T-GP50作BamHⅠ和XhoⅠ双酶切,得到与PCR扩增产物一致的插入片断,表明GP50编码基因具有一个长度为897 bp的完整开放阅读框,编码298个氨基酸,相对分子量33 000,与GenBank收录(编号为AY212945)的猪囊尾蚴GP50编码基因具有高度的同源性(99%)。结论:猪囊尾蚴GP50编码基因的克隆获得成功。

       

      Abstract: Objective:To set up a way of detecting cysticercosis quickly and accurately by use of recombinant protein,the GP50 gene was amplified from cysticercus cellulosae and cloned into pGEM-T vector.Methods:The specific primers were designed and synthesized.The DNA fragment encoding GP50 gene was amplified by PCR from cDNA of C.cellulosae.The PCR products were cloned into pGEM-T vector and recombinants were screened by PCR,enzyme digestion and sequencing.Results:The size of pGEM-T-GP50 digested with BamH Ⅰ and Xho Ⅰ was identical to the length of the PCR generated products.DNA sequencing indicated that the GP50 gene had an open reading frame of 897 bp,which encodes 298 amino acids with an approximate molecular weight of 33 000 and had high homology with the GP50 gene submitted to GenBank(AY212945).Conclusions:The findings suggested that GP50 gene had been successfully cloned.

       

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