乳腺肿瘤·转移抑制基因·BRMS1·基因表达_

乳腺癌转移抑制基因BRMS1真核表达载体的构建及鉴定

http://www.100md.com 《中国普通现代外科进展》 2006年第2期

乳腺肿瘤·转移抑制基因·BRMS1·基因表达,,乳腺肿瘤·转移抑制基因·BRMS1·基因表达,1材料与方法,2结果,3讨论,参考文献

     【摘要】目的：构建乳腺癌转移抑制基因(BRMS1)真核表达载体pcDNA3.1/mycBRMS1，为研究抑制恶性肿瘤转移的机制奠定基础。方法：设计含有Hind Ⅲ和XhoⅠ酶切位点的一对引物，应用逆转录聚合酶链式反应(RTPCR)技术从人乳腺癌细胞株MCF7中扩增到BRMS1的cDNA基因片断，经回收、纯化、酶切后，连接到质粒pcDNA3.1/mycHis(+)A上，鉴定正确后并转染人胚肾细胞HEK293进行Western blot验证其是否表达。结果：琼脂糖凝胶电泳及基因测序证实重组的pcDNA3.1/mycBRMS1 cDNA的碱基组成与GenBank中的人BRMS1 cDNA的基因片断组成相同。结论：成功构建了乳腺癌转移抑制基因BRMS1真核表达载体并测序鉴定。

    【关键词】乳腺肿瘤·转移抑制基因·BRMS1·基因表达

    Construction and identification of eukaryotic expression vector for human breast cancer metastasis suppressor1(BRMS1)

    TANG Lubing1,GAO Haidong1,TIAN Chunyan2,SUN Jingzhong1,LIU Huantao1,MA Rong1,WANG Tiantian1

    1Department of Breast Surgury,Qilu Hospital of Shandong University (Jinan 250012,

    China)

    2Institute of Radiation Medicine,Academy of Military Medical Sciences(Beijing

    100850,China)

    【ABSTRACT】Objective:To construct recombined eukaryotic expression vector pcDNA3.1/mycBRMS1cDNA successfully.This is useful for researching the mechanisms of metastatic suppression.Methods:To design a pair of primers,which two restriction site with HindⅢ and XhoⅠ,and BRMS1cDNA fragments were amplified from human breastcancer MCF7 by RTPCR.The product was extracted,purified and digested with Hind Ⅲ and Xho Ⅰ,then was inserted to PcDNA3.1/mycHis(+)A plasmid.The recombinants transfected into HEK293 cell and was identified by Western blot.Results:The base sequence of recombined pcDNA3.1/mycBRMS1cNDA was in accordance with human BRMS1 cDNA fragments by agarose gel electrophoresis and DNA sequence analysis.Conclusion:Eukaryotic expression vector for human breast cancer metastasis suppressor 1(pcDNA3.1/mycBRMS1)has been constructed successfuly and DNA sequence was analyzed. ......

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