| 人可溶性白细胞介素—6受体基因在昆虫细胞内的克隆与表达 |
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| 引用本文： | Chang BS,Li Y. 人可溶性白细胞介素—6受体基因在昆虫细胞内的克隆与表达[J]. 中国医学科学院学报, 2001, 23(1): 36-39 |
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| 作者姓名： | Chang BS Li Y |
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| 作者单位： | 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, |
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| 基金项目： | 中国医学科学院基金(971008)资助Supported by the Foundation of Chinese Academy of Medical Sciences |
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| 摘 要： | 目的：在昆虫细胞中表达人可溶性白细胞介素－6受体（sIL-6R)基因。方法：将人sIL-6R cDNA克隆至杆状病毒转移载体pAcGP67B中,再将重 组转移载体质粒与野生病毒AcNPV DNA共转染昆虫细胞sf9;经同源重组后,以终点稀释和斑点杂交法筛选重组杆状病毒;采用空斑分析法纯化重组的病毒,然后以纯化的重组病毒感染昆虫细胞Sf9。结果：斑点杂交实验证实,纯化得到的重组病毒含有人sIL-6R基因,SDS－PAGE结果显示,表达产物sIL-6R相对分子质量为47000左右,Western blot和受体配基结合实验表明,表达产物具有与其配基特异性地结合的能力,结论,杆状病毒－昆虫细胞能分泌表达sIL-6R,表达产物具有免疫活性和生物活性。
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| 关 键 词： | 可溶性白细胞介素－6受体 克隆 分泌表达 杆状病毒 |
| 修稿时间： | 2000-06-28 |
Gene cloning and expression of human soluble interleukin-6 receptor in insect cells |
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| Chang B S,Li Y. Gene cloning and expression of human soluble interleukin-6 receptor in insect cells[J]. Acta Academiae Medicinae Sinicae, 2001, 23(1): 36-39 |
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| Authors: | Chang B S Li Y |
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| Affiliation: | Department of Biotechnology, Institute of Medicinal Biotechnology, CAMS, PUMC, Beijing 100050, China. |
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| Abstract: | OBJECTIVE: To clone the human soluble interleukin-6 receptor(hsIL-6R) gene and expression in insect cell line. METHODS: The hsIL-6R gene was cloned into plasmid pAcGP67B. After co-transfection of recombinant plasmid and wild type AcNPV DNA, the rAcNPV was confirmed by the end point dilution assay and dot blot. Then it was purified by plaque assay. RESULTS: SDS-PAGE showed molecular weight of the expressed product was about 47,000. The expressed recombinant protein was confirmed to be specific and capable of binding its ligand IL-6 by Western blot and ligand-receptor binding assays. CONCLUSIONS: Secretory expression of hsIL-6R gene in baculovirus expression system was indicated. The expressed product had immunological and biological activities. |
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| Keywords: | soluble human interleukin- 6 receptor gene cloning and expression baculovirus |
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