| IL15/PAP融合蛋白基因克隆及其原核与植物表达载体的构建 |
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| 引用本文： | 陈其新,陈凌娜,陈正华.IL15/PAP融合蛋白基因克隆及其原核与植物表达载体的构建[J].河南农业大学学报,2006,40(3):301-306. |
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| 作者姓名： | 陈其新 陈凌娜 陈正华 |
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| 作者单位： | 中国科学院西北高原生物所，新疆农业大学，甘肃亚盛集团博士后科研工作站 青海西宁830000，河南农业大学，河南郑州450002，新疆乌鲁木齐810000，甘肃兰州730070 |
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| 摘 要： | 将IL15和PAP基因通过一甘氨酸接头(Gly4Ser)3连接在一起,构建原核和植物表达载体,以期表达具有导向杀伤活性的融合蛋白.在引物设计中,通过引物在IL15基因下游引入(Gly4Ser)3的编码序列和BamHI位点,然后采取分步克隆及PCR、酶切、连接等一系列分子克隆方法,将IL15和PAP基因融合在一起,构建融合蛋白的原核与植物表达载体.经PCR和NcoI/SalI双酶切鉴定及测序,证实原核表达载体pET32a-IL15/PAP及植物表达载体pB-I P中均插入了正确的融合基因序列.
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| 关 键 词： | IL15/PAP融合蛋白 表达载体 构建 |
| 文章编号： | 1000-2340（2006）03-0301-06 |
| 收稿时间： | 2005-10-20 |
| 修稿时间： | 2005年10月20 |
Gene Cloning and Construction of Prokaryotic and Plant Expression Vectors of IL15/PAP Fusion Protein |
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| CHEN Qi-xin, CHEN Ling-na, CHEN Zheng-hua.Gene Cloning and Construction of Prokaryotic and Plant Expression Vectors of IL15/PAP Fusion Protein[J].Journal of Henan Agricultural University,2006,40(3):301-306. |
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| Authors: | CHEN Qi-xin CHEN Ling-na CHEN Zheng-hua |
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| Institution: | 1. Northwest Institute of Plateau Biology, CAS, Xining 830000, China ; 2. Postdoctoral Research Station of Yasheng Group Co., Lanzhou 730070, China; 3. Xinjiang Agricultural University, Uramqi 810000, China ; 4. Henan Agricultural University, Zhengzhou 450002, China |
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| Abstract: | To link IL15 gene and PAP gene by a mediate sequence(Gly-4Ser)-3 and express a new kind of fusion protein with target killing effects on the tumor cells overexpressing IL15R in E.coli and plants,a DNA linker consisting of(Gly-4Ser)-3 coding sequence and Bam HI restriction site was introduced into the downstream of IL15 gene,then the vectors for expressing the fusion protein in E.coli and plants were constructed by step-by-step cloning and PCR,enzyme digestion and linkage techniques.The PCR,restriction analysis with Nco I/Sal I and sequencing claimed that the fusion gene of interest was inserted into the prokaryotic expression vector pET32a-IL15/PAP and the plant expression vector pBIP respectively. |
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| Keywords: | IL15/PAP fusion protein expression vector construction |
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