基因克隆__(期刊论文)

编码表皮生长因子受体Ⅲ型突变体PEP3表位的cDNA的克隆

http://www.100md.com 《西安交通大学学报》 2006年第2期

基因克隆,,表皮生长因子受体Ⅲ型突变体；PEP3表位；cDNA；基因克隆,1材料与方法,2结果,3讨论,参考文献：

     作者简介：段小艺(1974)，女(汉族)，讲师，博士研究生. 研究方向：肿瘤核医学.

    摘要：目的克隆出针对表皮生长因子受体Ⅲ型突变体(EGFRvⅢ)PEP3表位的cDNA片段，为高表达EGFRvⅢ肿瘤的免疫治疗奠定基础。方法根据PEP3的碱基序列设计出有12对互补碱基的正负引物，正负引物互为模板进行PCR扩增，制备出两端含酶切位点的PEP3 cDNA片段，再将扩增的PCR产物亚克隆于载体pGEMT Easy构建重组质粒pGEMT Easy/PEP3。结果经限制性内切酶酶切鉴定和DNA测序证实，编码PEP3的cDNA片段被成功扩增并亚克隆于载体pGEMT Easy中。结论成功克隆了编码表皮生长因子受体Ⅲ型突变体PEP3表位的cDNA片段。

    关键词：表皮生长因子受体Ⅲ型突变体；PEP3表位；cDNA；基因克隆

    Clone of cDNA encoding PEP3 epitope of epidermal growth factor receptor type Ⅲ mutantDuan Xiaoyi, Wang Jiansheng, Guo Youmin, Liu Min,Zhou Bin, Wang Quanying, Yang Guangxiao

    (1. Department of Nuclear Medicine;

    2. Department of Oncosurgery, First Affiliated Hospital,Medical School of Xian Jiaotong University, Xian 710061;

    3. Image Centre, SecondAffiliated Hospital, Medical School of Xian Jiaotong University, Xian 710004;

    4. Xian Huaguang Biological Engineering Company, Xian 710025, China)

    ABSTRACT: Objective Through cloning the cDNA fragment encoding PEP3 epitope of epidermal growth factor receptor type Ⅲ mutant(EGFRvⅢ) to establish the base for immunotherapy of carcinoma with high EGFRvⅢ expressing. Methods Designing the primer according to the base sequence of PEP3, cDNA was synthesized and amplificated by PCR. Then, the PCR product was subcloned into vector pGEMT Easy so as to construct recombinant plasmid pGEMT Easy/ PEP3. Results Zymocutting restriction enzyme identification and DNA sequencing conformed that cDNA encoding PEP3 was amplified and subcoloned into pGEMT Easy successfully. Conclusion cDNA encoding PEP3 epitope of EGFRvⅢ was coloned successfully. ......

百拇医药网 http://www.100md.com/html/DirDu/2006/06/10/00/78/13.htm

您现在查看是摘要页，全文长 8619 字符。