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| 人CD96基因胞外区的原核表达及抗血清制备 | |
| 引用本文： | 曾建明,刘飞,谭坪海,王丽娜,李沫,陈中华,李松,龙一飞,李有强,陈茶.人CD96基因胞外区的原核表达及抗血清制备[J].南方医科大学学报,2011,31(7):1232-1235. |
| 作者姓名： | 曾建明 刘飞 谭坪海 王丽娜 李沫 陈中华 李松 龙一飞 李有强 陈茶 |
| 作者单位： | 广东省中医院检验科;南方医科大学南方医院检验科;广东医学院检验学院; |
| 基金项目： | 广东省医学科学技术研究基金(B2008066) |
| 摘 要： | 摘要：目的原核表达并纯化人CD96胞外区蛋白，免疫家兔制备抗体。方法构建pET32-CD96重组质粒，转化大肠埃希 菌BL21(DE3)，以IPTG诱导表达，Ni2+-NTA树脂纯化蛋白并免疫家兔，以间接ELISA检测抗体效价，Western blot鉴定抗 体特异性。结果质粒pET32-CD96经测序鉴定，通过SDS电泳显示能在BL21(DE3)中高效表达，相对分子质量约37 000； Western blot实验结果显示制备的多克隆抗体可与HL-60细胞提取蛋白及原核表达的人CD96蛋白胞外区特异性结合。 结论成功构建并表达人CD96胞外区蛋白，免疫家兔获得高滴度抗血清，为后续研究奠定了基础。 |
| 关 键 词： | 人CD96基因 原核表达 白血病干细胞 多克隆抗体 |
Human CD96 gene cloning,expression and identification |
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| ZENG Jian-ming,LIU Fei,TAN Ping-hai,WANG Li-na,LI Mo,CHEN Zhong-hua,Li Song,LONG Yi-fei,LI You-qiang,CHEN Cha.Human CD96 gene cloning,expression and identification[J].Journal of Southern Medical University,2011,31(7):1232-1235. | |
| Authors: | ZENG Jian-ming LIU Fei TAN Ping-hai WANG Li-na LI Mo CHEN Zhong-hua Li Song LONG Yi-fei LI You-qiang CHEN Cha |
| Institution: | ZENG Jian-ming 1,LIU Fei 2,TAN Ping-hai 3,WANG Li-na 1,LI Mo 1,CHEN Zhong-hua 1,Li Song 1,LONG Yi-fei 1,LI You-qiang 1,CHEN Cha 1 1 Department of Laboratory Medicine,Guangdong Provincial Hospital of Traditional Chinese Medicine,Guangzhou 510120,China,2 Department of Laboratory Medicine,Nanfang Hospital Southern Medical Univesity,Guangzhou 510515,3 Institute of Laboratory Medicine,Guangdong Medical College,Dongguan 523808 |
| Abstract: | Objective To construct and express human CD96 gene outer membrane domain(hCD96om) in prokaryotic cells and prepare rabbit polyclonal antibody of hCD96om.Methods hCD96om was amplified by RT-PCR from the peripheral blood of patients with acute myeloid leukemia and inserted into prokaryotic expression vector pET32a(+) to construct the recombinant plasmid pET32-CD96.The expression of hCD96om was induced by IPTG in BL21(DE3) cells,and the expression product was identified by Western blotting.The anti-hCD96 polyc... |
| Keywords: | human CD96 gene prokaryotic expression leukemic stem cell polyclonal antibodies |
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