[1]王颖莹,陈纯海,裴莉萍,等.2型大麻受体的克隆及真核表达体系的构建[J].第三军医大学学报,2007,29(11):1003-1005.
 WANG Ying-ying,CHEN Chun-hai,PEI Li-ping,et al.Clone and eukaryotic expression of human cannabinoid receptor 2 gene[J].J Third Mil Med Univ,2007,29(11):1003-1005.
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2型大麻受体的克隆及真核表达体系的构建
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
29卷
期数:
2007年第11期
页码:
1003-1005
栏目:
论著
出版日期:
2007-06-15

文章信息/Info

Title:
Clone and eukaryotic expression of human cannabinoid receptor 2 gene
作者:
王颖莹陈纯海裴莉萍张伟 Norbert E. Kaminski
第三军医大学:西南医院检验科,军事预防医学院劳动卫生学教研室;美国密西根州立大学药理与毒理系
Author(s):
WANG Ying-ying CHEN Chun-hai PEI Li-ping ZHANG Wei Norbert E. Kaminski
Department of Clinical Laboratory, Southwest Hospital, Department of Occupational Hygiene, College of Preventive Medicine, Third Military Medical University, Chongqing 400038, China; Department of Pharmacology and Toxicology, Michigan State University, East Lansing 48824, USA
关键词:
2型大麻受体基因克隆真核表达
Keywords:
CB2 gene cloning eukaryotic expression
分类号:
R392.2;R394-33
文献标志码:
A
摘要:
目的    克隆人2型大麻受体(hCB2)基因,并构建相应的稳定表达细胞株。    方法    利用人T淋巴细胞株HPB-ALL的总RNA,以RT-PCR及酶切、连接等分子生物学方法克隆hCB2基因于质粒载体pFlag-CMV-3内,以HEK293细胞构建稳定表达细胞株,以Western blot方法利用蛋白标记Flag的单克隆抗体(Anti-Flag)和CB2多克隆抗体(Anti-CB2)同时检测质粒的表达。    结果    酶切及测序结果表明hCB2基因克隆成功,Western blot检测结果表明Anti-Flag和Anti-CB2都能检测Flag-hCB2蛋白。    结论    成功克隆hCB2基因,并获得稳定表达细胞株,同时证实Anti-CB2多克隆抗体有效。
Abstract:
Objective    To clone human cannabinoid receptor 2 gene and establish stable cell line which constantly expresses hCB2 protein.     Methods    Full length of hCB2 cDNA was obtained by RT-PCR with total RNA isolated from human T cell line HPB-ALL, and then constructed into plasmid vector pFlag-CMV-3. The stable cell line was established with HEK293 cells. The expression of Flag-CB2 was analyzed by Western blotting with monoclonal Anti-Flag and multiclonal Anti-CB2.     Results    pFLag-hCB2 was confirmed containing proper hCB2 gene with diagnostic digestion and sequencing, and the expression products were detected by Anti-Flag and Anti-CB2 parallelly.     Conclusion    Gene of hCB2 has been cloned and constantly expressed in HEK293 cells. The multiclonal Anti-CB2 is proved efficient in detecting CB2 protein epitope.

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更新日期/Last Update: 2008-10-23