乳腺生长抑制因子mammastatin表位抗原的克隆表达及初步血清学测定  

Breast growth inhibitor mammastatin:its cloning expression of epitope and primary serum detection

作　　者：黄焰[1] 宋晓国[2] 张贺秋[2] 王国华[2] 王刚乐[1] 官奇 尉承泽[1] 

机构地区：[1]军事医学科学院附属医院,北京100071 [2]军事医学科学院基础医学研究所,北京100850

出　　处：《军事医学科学院院刊》2008年第2期151-153,198,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：全军“十一五”面上课题(06MA352)

摘　　要：目的:获得重组乳腺生长抑制因子(mammastatin)表位抗原蛋白,制备抗血清,以测定正常人及乳腺癌患者血清中mammastatin存在水平。方法:用BioSun生物软件对mammastatin氨基酸序列进行分析,确定表位序列,PCR逐步延伸法合成表位基因,构建pBVIL1-mammastatin和pBVIL6-mammastatin表达质粒,转化于大肠杆菌HB101中进行表达,经纯化后免疫大耳白兔,制备抗血清,用双抗体夹心法检测乳腺癌及正常人的血清。结果:重组mammastatin抗原在大肠杆菌中获得了高效表达,并获得了兔多抗血清。用双抗体夹心法检测,显示95例正常对照血清mammastatin表达水平显著高于95例乳腺癌(平均D值,正常组0.261,乳腺癌组0.135,Wilcoxon秩和检验P=0.000<0.01,差异显著)。结论:正常人血清中乳腺生长抑制因子mammastatin存在水平显著高于乳腺癌患者。Objective： To prepare the recombinant epitope protein and antiserum of mammastatin to determine mammastatin levels in the sera of normal people and breast cancer patients. Methods： The amino acid sequence of mammastatin was analyzed by BioSun to determine the epitope sequence. The epitope gene was synthesized by PCR. The expression vectors of pBVIL1-mammastatin and pBVIL6-mammastatin were constructed and transformed into E. coli HB101 for expression. Rabbits were immunized with the purified recombinant protein to prepare antiserum for the detection of mammastatin in the sera of normal people and breast cancer patients by ELISA. Results：Recombinant mammastatin antigen was effectively expressed in E. coli and the antiserum was prepared. The sera of 95 normal people and 95 breast cancer patients were analyzed by ELISA. The expression level of mammastatin in normal people（average D value 0. 261 ） was significantly higher than that in breast cancer patients （ average D value 0. 135, Wileoxon rank test P = 0.000 〈 0.01 ）. Conclusion： Breast growth inhibitor mammastatin has a higher level in the serum of normal people than in breast eancer patients.

关 键 词：乳腺肿瘤 克隆表达 乳腺生长抑制因子 mammastatin 表位 质粒 血清学 

分 类 号：R392[医药卫生—免疫学]