| 小鼠T细胞活化衔接因子基因真核表达载体的构建 |
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| 引用本文： | 任涟萍,郭雪君. 小鼠T细胞活化衔接因子基因真核表达载体的构建[J]. 上海交通大学学报(医学版), 2007, 27(2): 168-170 |
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| 作者姓名： | 任涟萍 郭雪君 |
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| 作者单位： | 上海交通大学,医学院新华医院呼吸内科,上海,200092;上海交通大学,医学院新华医院呼吸内科,上海,200092 |
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| 摘 要： | 目的构建小鼠T细胞活化衔接因子(LAT)基因的真核表达载体。方法以小鼠单个核细胞的cDNA为模板,采用PCR扩增LAT基因编码区的全部序列,克隆入真核细胞表达载体pCMV-HA中;测序鉴定目的基因并转染哮喘小鼠脾脏淋巴细胞。结果PCR扩增的特异性片段长度为729 bp,并以此构建重组质粒pCMV-HA-LAT。质粒测序结果与GenBank中的小鼠LAT基因cDNA序列一致;转染哮喘小鼠淋巴细胞后可检测到LAT蛋白的表达。结论成功构建了pCMV-HA-LAT重组真核表达载体。
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| 关 键 词： | LAT 基因克隆 真核表达载体 哮喘 |
| 文章编号： | 0258-5898(2007)02-0168-03 |
| 修稿时间： | 2006-05-26 |
Construction of linker for activation of T cells gene eukaryotic expression vector in mouse |
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| REN Lian-ping,GUO Xue-jun. Construction of linker for activation of T cells gene eukaryotic expression vector in mouse[J]. Journal of Shanghai Jiaotong University:Medical Science, 2007, 27(2): 168-170 |
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| Authors: | REN Lian-ping GUO Xue-jun |
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| Affiliation: | Department of Respiratory Medicine, Xinhua Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200092, China |
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| Abstract: | Objective To construct the eukaryotic expression vector of linker for activation of T cells(LAT) gene in mouse. MethodsLAT cDNA of mouse mononuclear cells was amplified with polymerase chain reaction.The fragment was inserted into the eukaryotic expression vector of pCMV-HA plasmid.The recombinant plasmid was verified by DNA sequencing and used to transfect lymphocytes of the asthmatic model. Results The length of amplified fragment was 729 bp.The sequence of LAT gene was confirmed by Genbank.The LAT protein could be detected in the lymphocytes of asthmatic model.Conclusion The recombinant eukaryotic expression vector pCMV-HA-LAT can be successfully constructed. |
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| Keywords: | LAT |
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