| p16基因的原核表达及抗P16抗血清的制备 |
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| 引用本文： | 覃林花,黄长晖,博继梁. p16基因的原核表达及抗P16抗血清的制备[J]. 生物技术通讯, 1998, 0(3) |
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| 作者姓名： | 覃林花 黄长晖 博继梁 |
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| 作者单位： | 第二军医大学医学分子遗传学军队重点实验室，第二军医大学医学分子遗传学军队重点实验室，第二军医大学医学分子遗传学军队重点实验室 上海 200043 诲军411医院消化实验室，上海 200081，上海 200043，上海 200043 |
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| 基金项目： | 国家自然科学基金(项目号 39670400) |
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| 摘 要： | 采用基因重租技术,将野生型p16 cDNA通过中间载体pVL1392最后载入表达载体pGEX-5T,IPTG诱导重组质粒转化的大肠杆菌,蛋白质印迹证实42×10~3的融合蛋白GST-P16的表达。表达了GST-P16的重组菌经加热破菌后行SDS-PAGE,将GST-P16蛋白条带切下后经反复冻融于皮内多点注射免疫家兔。所收获的兔血清经蛋白质印迹法检测到抗P16抗体滴度为1:625。结果表明通过pGEX-5T融合蛋白表达系统能方便地提供制备抗P16抗血清的免疫原,该免疫原未经纯化并未影响抗P16抗血清的产生。
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| 关 键 词： | p16基因(CDKN2) 原核表达 蛋白免疫 |
Expression of p16 as fusion protein in E. coli and production of antisera to GST-P16 |
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| Qin Linhua,Huang Changhui,Fu Jiliang. Expression of p16 as fusion protein in E. coli and production of antisera to GST-P16[J]. Letters in Biotechnology, 1998, 0(3) |
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| Authors: | Qin Linhua Huang Changhui Fu Jiliang |
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| Abstract: | p16 cDNA was cloned into expression vector pGEX-5T and transformed E. coli TG1. After 3 hours induced by IPTG, the expression of recombinant 42 kD GST-pl6 was confirmed by SDS-PAGE and Western blotting. For producing antibody to p16. the recombinant GST-p16 fusion protern belt was cut out from SDS-PAGE gel and emulsified with adjuvant to be as antigen to inoculate rabbits. After the rabbits were subcutaneous injected with antigen protein about 0. 5 mg in 0 day, 28 day and 56 day, the titer of anti-p16 antisera reached to 1 : 625 when examined by Western blotting. |
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| Keywords: | p16 gene (CDKN2) prokaryotic expression protein immunization |
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