| 人杀伤抑制性受体CD158bcDNA的克隆及其在大肠杆菌中的表达 |
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| 引用本文： | 曹东林,周思朗,白英明,何肖娟. 人杀伤抑制性受体CD158bcDNA的克隆及其在大肠杆菌中的表达[J]. 第一军医大学分校学报, 2004, 27(1): 10-12 |
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| 作者姓名： | 曹东林 周思朗 白英明 何肖娟 |
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| 作者单位： | 第一军医大学分校生化教研室,广东省广州市,510315;海军421医院老干病房,广东省广州市,510318 |
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| 基金项目： | 国家自然科学基金，广东省自然科学基金 |
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| 摘 要： | 目的:CD158b基因克隆与表达.方法:采用RT-PCR技术,从正常人外周血单个核细胞中扩增编码CD158b的cDNA,经酶切后将其克隆于pMBP-c表达质粒上,酶切和测序鉴定.构建的高效表达克隆子经IPTG诱导后,表达的重组蛋白经SDS-PAGE电泳分析.结果:RT-PCR获得了预期大小的cDNA,DNA测序结果与GenBank登记的人CD158b编码序列一致.pMBP-c-CD158b表达质粒酶切后结果与预期相符.表达的重组蛋白经SDS-PAGE电泳分析,得到分子量为58 kD的蛋白质,与理论值一致,所获重组蛋白占菌体总蛋白25%.结论:获得了CD158b基因,并成功在大肠杆菌中表达,所获重组蛋白为进一步研究CD158的功能打下了基础.
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| 关 键 词： | CD158b基因 逆转录聚合酶链式反应 表达质粒 |
| 文章编号： | 1009-1793(2004)01-0010-03 |
| 修稿时间： | 2003-10-22 |
Cloning of human natural killer cell inhibitory receptor CD158b cDNA and its expression in E.coli |
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| CAO Dong lin,ZHOU Si lang,BAI Ying ming,et al.. Cloning of human natural killer cell inhibitory receptor CD158b cDNA and its expression in E.coli[J]. Journal of Branch Campus of the First Military Medical University, 2004, 27(1): 10-12 |
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| Authors: | CAO Dong lin ZHOU Si lang BAI Ying ming et al. |
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| Affiliation: | CAO Dong lin,ZHOU Si lang,BAI Ying ming,et al . |
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| Abstract: | Objective To clone and express the CD158b gene.Methods The gene which encodes matural killer cell inhibitory receptor was amplified by RT-PCR from peripheral blood mononuclear cells(PBMCs). Recombinant expression vector was constructed and sequenced after enzyme digestion. Results (1)Prospective amplified cDNA was obtained and the base sequence was identical with that recorded in GenBank after DNA sequencing. (2)Identification of pMBP-c-CD158b expression vector showed prospective result after the enzyme digestion analysis. (3)The size of recombinant protein was also coincident with the anticipotive aim and its proportion was 25% of the total protein. Conclusion CD158b cDNA was obtained by RT-PCR and successfully expressed in E.coli . The recombinant protein could provide an essential preparation for studying its functions. |
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| Keywords: | CD158b gene RT-PCR expression vector |
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