| 十二指肠钩虫金属蛋白酶组织抑制剂同源物Ad-TIMPL-1基因克隆及其原核表达 |
| |
| 引用本文： | 邓莉,邵正,许琴英,胡晶晶,彭礼飞.十二指肠钩虫金属蛋白酶组织抑制剂同源物Ad-TIMPL-1基因克隆及其原核表达[J].热带医学杂志,2011,11(8):866-869,874. |
| |
| 作者姓名： | 邓莉 邵正 许琴英 胡晶晶 彭礼飞 |
| |
| 作者单位： | 广东医学院寄生虫学教研室,广东湛江,524023 |
| |
| 基金项目： | 广东省自然科学基金(04011381) |
| |
| 摘 要： | 目的克隆十二指肠钩虫金属蛋白酶组织抑制剂同源物(TIMPL)Ad-TIMPL-1基因并进行原核表达。方法分别用3’RACE及RT-PCR技术从十二指肠钩虫成虫cDNA中扩增编码Ad-TIMPL-1的cDNA3’末端及5’末端序列;序列经拼接后进行初步生物信息学分析;将Ad-TIMPL-1成熟肽编码序列克隆至原核表达载体pET-32a,构建重组表达质粒;重组质粒转化至大肠杆菌BL21(DE3)后,用IPTG诱导表达,SDS-PAGE分析表达情况。结果成功克隆获得了编码Ad-TIMPL-1的全长cDNA序列并登记到GenBank(accession no.EF495071);Ad-TIMPL-1完整阅读框为396bp,编码132个氨基酸残基组成的蛋白,含16个氨基酸残基组成的信号肽;成功构建了原核表达重组质粒pET-32a/Ad-TIMPL-1,并在大肠杆菌中得到了表达。结论本研究首先从十二指肠钩虫中克隆到了Ad-TIMPL-1基因并进行了原核表达,为进一步研究其生物学功能奠定了基础。
|
| 关 键 词： | 十二指肠钩虫 金属蛋白酶组织抑制剂 克隆 原核表达 |
Cloning and prokaryotic expression of Ad-TIMPL-1,a tissue inhibitor of metalloproteinase homologue from Ancylostoma duodenale |
| |
| DENG Li,SHAO Zheng,XU Qin-ying,HU Jing-jing,PENG Li-fei.Cloning and prokaryotic expression of Ad-TIMPL-1,a tissue inhibitor of metalloproteinase homologue from Ancylostoma duodenale[J].Journal Of Tropical Medicine,2011,11(8):866-869,874. |
| |
| Authors: | DENG Li SHAO Zheng XU Qin-ying HU Jing-jing PENG Li-fei |
| |
| Institution: | DENG Li,SHAO Zheng,XU Qin-ying,HU Jing-jing,PENG Li-fei (Department of Parasitology,Guangdong Medical College,Guangdong,Zhanjiang 524023,China) |
| |
| Abstract: | Objective To clone and express the tissue inhibitor of metalloproteinase homologue(TIMP like,TIMPL),Ad-TIMPL-1,from the hookworm Ancylostoma duodenale.Methods The 5' and 3' end of cDNA encoding Ad-TIMPL-1 were amplified by 3'RACE and RT-PCR from the cDNA of adult A.duodenale,respectively.The nucleotide sequence encoding mature Ad-TIMPL-1 was ligated into pET32a to construct the recombinant plasmid pET32a/Ad-TIMPL-1.The recombinant plasmid was transformed into E.coli BL21(DE3),and then expressed by inducing ... |
| |
| Keywords: | Ancylostoma duodenale Ad-TIMPL-1 cloning prokaryotic expression |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
