肿瘤转移抑制基因nm23-H1和nm23-H2特异性片段的体外扩增、克隆和鉴定  

Amplification, Clone and Identification of the Specific Fragments of Tumor MetastasisSuppressor Gene nm23H 1 and nm23H 2

作　　者：李昌龙[1,2] 卢勇[1,2] 王若菡[1,2] 陈红英 陈俊杰[1,2] 

机构地区：[1]华西医科大学生物化学教研室 [2]卫生部口腔生物医学工程重点实验室

出　　处：《华西口腔医学杂志》1997年第4期322-324,I013,共3页West China Journal of Stomatology

基　　金：国家自然科学基金

摘　　要：采用聚合酶链反应（ＰＣＲ），从人基因组ＤＮＡ中扩增出ｎｍ２３－Ｈ１（１８５ｂｐ）和ｎｍ２３－Ｈ２（１４５ｂｐ）特异性ＤＮＡ片段，与载体ｐＧＥＭ－３ｚｆ（＋）平端连接，重组质粒分别转化宿主菌ＪＭ１０９，在含Ｘ－ｇａｌ和ＩＰＴＧ的平板上直接筛选阳性克隆，限制性内切酶谱分析及ＰＣＲ扩增鉴定，确定ｐＧＥＭ－Ｈ１和ｐＧＥＭ－Ｈ２重组体。为ｎｍ２３基因与肿瘤转移研究打下基础。ァ　series of DNA primers specific for the specific fragments of nm23H1 and nm23H2 were designed and synthesized.The specific fragments of nm23H1 (185bp) and nm23H2 (145bp) were amplified from human blood DNA by using poly merase chain reaction (PCR).The recovered PCR products were treated with Klenow fragment,inserted into pGEM 3 zf (+)vector with bluntend ligation,and then transformed into competent cell J M109.The positive colonies were directly iden tified by colour screening on indicator plates.The recombinant plasmids were digested by Alu Iand identified by PCR.There sults showed that the authors had obtained the specific fragments of nm23H1and nm23H2 respectively,and these specific fragm ents could be used for study the expression of nm23H1 and nm23H2 seperately.

关 键 词：抑癌基因 体外扩增 克隆 肿瘤转移 

分 类 号：R73-37[医药卫生—肿瘤]