聚合酶链反应用于人巨细胞病毒B基因编码区段的体外扩增  

Application of Polymerase Chain Reaction in Amplification of Human Cytomegalovirus B Gene Code Region

作　　者：吴均[1] 陈俊杰[2] 唐泽媛[1] 顾建仁[3] 陈渊卿[3] 李岱宗[3] 

机构地区：[1]华西医大儿科学教研室 [2]华西医大生物化学教研室 [3]上海市肿瘤研究所生物化学及分子生物学研究室

出　　处：《华西医科大学学报》1991年第2期117-119,共3页Journal of West China University of Medical Sciences

基　　金：高等学校博士学科点专项基金

摘　　要：作者采用聚合酶链反应(PCR)成功地从含人巨细胞病毒B基因的重组质粒pBH_2DNA中扩增及分离了B基因编码区段及其产物糖蛋白52kd抗原编码区段。0.2μg的模板DNA经过30次循环,目的基因片段的扩增量达到10μg,足以用于酶谱分析及克隆的构建。A nucleic acid amplifi-cation procedure, the polymerase chainreaction (PCR), has been used to amp-lify the human cytomegalovirus (HCMV)B gene code region, and its glycoprotein52 kd atigentic domain. The primersused in the PCR assay were based on aconserved region of the HCMV B genesequence. By using primer 1 (5′-AAAGAATTCATGGAATCCAGGATCT G-3′up-stream nucleotides 157 to 2877), primer 2(5′-AAAGAATTCATGAACGTGAAGGA-ATCG-3′,upstream nucleotides 1846 to2877), and primer 4 (5′-ATAAAGCTTAATCAGACGTTCTCTTCTTC-3′, downstreamnucleotides 157 to 2877 and 1846 to 2877) ,the HCMV B gene code region sequenceand its glycoprotein 52 kd antigentic do-main sequence were amplified from therecombintal plasmid pBH1 DNA contain-ing the HCMV B gene. Amplificationcycles consisted of denaturation at 92℃for 1 min, annaling at 55℃ for 1 min,and extension at 72℃ for 1.5 min. Thecycles were carried out 30 times in threedifferent water baths, respectively. Afterthe last extension, 10 μl of each reactionmixture was removed and subjected toelectrophoresis on 1.2% agarose gel. Ten μgof PCR products were obtained from0.2μg of template DNA after the 30times cycles, and were enough for beingused in the restriction site analysis andthe construction of clones.

关 键 词：聚合酶链反应 人巨细胞病毒 B基因 

分 类 号：R394.8[医药卫生—医学遗传学]