| CTLA—4在大肠杆菌中的克隆和表达 |
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| 引用本文： | Wang CJ,Li Y. CTLA—4在大肠杆菌中的克隆和表达[J]. 中国医学科学院学报, 2001, 23(2): 154-157 |
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| 作者姓名： | Wang CJ Li Y |
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| 作者单位： | 1. 中国医学科学院中国协和医科大学医药生物技术研究所生物工程室,
2. Department of Bioengineering, Institute of Medicinal Biotechnology, CAMS and PUMC, |
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| 摘 要： | 目的 在原核表达系统大肠杆菌中表达具有生物活性的人毒性T淋巴细胞相关蛋白－4可溶性部分（human soluble CTLA-4)。方法 PCR扩增获得CTLA－4编码基因,利用表达载体pGEX－2T构建重组质粒2TC,在所获DNA片段与文献报道CTLA－4序列一致。在重组大肠杆菌中0．10mmol/L IPTG诱导4h可大量生成相对分子质量40000的GST－CTLA4融合蛋白。Westen blot分析表明原核表达产物CTLA－4具抗原抗体结合活性。结论 hsCTLA－4可在原核表达系统中表达并具有免疫活性。
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| 关 键 词： | CTLA－4 基因克隆 原核表达 大肠杆菌 |
| 修稿时间： | 2000-09-25 |
Gene cloning and expression of CTLA-4 in E. coli |
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| Wang C J,Li Y. Gene cloning and expression of CTLA-4 in E. coli[J]. Acta Academiae Medicinae Sinicae, 2001, 23(2): 154-157 |
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| Authors: | Wang C J Li Y |
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| Affiliation: | Department of Bioengineering, Institute of Medicinal Biotechnology, CAMS, PUMC, Beijing 100050, China. |
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| Abstract: | OBJECTIVE: To express hsCTLA-4 in E. coli. METHODS: The hsCTLA-4 gene was obtained by PCR amplification from pE plasmid which contains CTLA-4 gene and was inserted into the expression vector pGEX-2T. The recombination strain was induced by IPTG with different concentrations and time. RESULTS: The sequence of PCR amplified DNA fragments was identical with the reported CTLA-4 gene. SDS-PAGE and Western blot showed that 0.10 mmol/L IPTG can induce higher production of the fusion protein with molecular weight 40,000 after addition of IPTG 4 hours and GST-CTLA4 had immunological activity. CONCLUSIONS: hsCTLA-4 can be expressed in soluble form high efficiency. |
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| Keywords: | CTLA-4 gene cloning prokaryotic expression |
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