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| 近交五指山小型猪SLA-DQA基因的克隆及在大肠杆菌的原核表达 | |
| 引用本文： | 王晓东,牟玉莲,张莉,杨树林,罗玉柱,冯书堂.近交五指山小型猪SLA-DQA基因的克隆及在大肠杆菌的原核表达[J].甘肃农业大学学报,2006,41(4):23-26. |
| 作者姓名： | 王晓东 牟玉莲 张莉 杨树林 罗玉柱 冯书堂 |
| 作者单位： | 1. 甘肃农业大学动物科学技术学院,甘肃,兰州,730070;中国农业科学院畜牧研究所,北京,100094 2. 中国农业科学院畜牧研究所,北京,100094 3. 甘肃农业大学动物科学技术学院,甘肃,兰州,730070 |
| 基金项目： | 国家高技术研究发展计划(863计划) , 科技基础性工作项目 , 中央级科研院所社会公益研究专项基金 |
| 摘 要： | 采用RT-PCR方法扩增五指山猪SLA-DQA基因cDNA,克隆入测序载体,测序结果与Gene Bank(登陆号:AY243104)的序列相同,大小为768 bp,该基因编码255个氨基酸残基和一个终止密码,预计蛋白分子量为28 kD.将这个基因克隆到原核表达载体PET-28a(+)中,经酶切、PCR扩增和测序分析确证其正确,插入到表达载体中,阅读框是正确的,从而构建成重组表达载体PET-28a(+)-DQA.重组质粒转化至宿主菌BL21(DE3)中,用IPTG进行诱导表达,对表达的蛋白用SDS-PAGE电泳分析,结果表明得到了与预计分子量相同的蛋白. |
| 关 键 词： | SLA-DQA基因 cDNA 克隆 原核表达 |
| 文章编号： | 1003-4315（2006）04-0023-04 |
| 收稿时间： | 2006-02-28 |
| 修稿时间： | 2006-02-28 |
Cloning and prokaryotic expression of SLA- DQA gene of inbred miniature Wuzhishan pig |
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| WANG Xiao-dong,MOU Yu-lian,ZHANG Li,YANG Shu-lin,LUO Yu-zhu,FENG Shu-tang.Cloning and prokaryotic expression of SLA- DQA gene of inbred miniature Wuzhishan pig[J].Journal of Gansu Agricultural University,2006,41(4):23-26. | |
| Authors: | WANG Xiao-dong MOU Yu-lian ZHANG Li YANG Shu-lin LUO Yu-zhu FENG Shu-tang |
| Institution: | 1. College of Animal Science and Technology, Gansu Agricultural University,Lanzhou 730070; 2. Institute of Animal Sciences,Chinese Academy of Agricultural Sciences,Beijing 100094,China |
| Abstract: | The cDNA of SLA-DQA gene of inbred miniature Wuzhishan pig was amplified by RT-PCR,and was cloned into PGEM-T vector for sequencing and analysis.The sequence of SLA-DQA cDNA showed no difference with the known sequence in the Gene bank.The SLA-DQA cDNA was cloned into expression vector PET-28a( ),after enzyme digestion and sequence analysis,it was testified that the open reading frame was right,then the recombinant expression vector PET-28a( )/SLA-DQA was constructed.The PET-28a( )/SLA-DQA was transformed into host bacterium,BL21(DE3),and was induced to express by IPTG,the purpose protein were identified by SDS-PAGE.The results indicated that the target proteins expressed by Escherichia coli were the same as expected. |
| Keywords: | SLA-DQA gene cDNA clone prokaryotic expression |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |