登革2型病毒NGC株基因组亚克隆的构建及其鉴定  

Construction and identification of genomic cDNA subclones of dengue 2 virus NGC strain

作　　者：贡树基[1] 曹虹[1] 赵卫[2] 张文炳[3] 周浩[3] 陈丽丹[3] 

机构地区：[1]南方医科大学公共卫生与热带医学学院微生物学系,广东广州510515 [2]南方医科大学公共卫生与热带医学学院病毒研究所,广东广州510515 [3]南方医科大学公共卫生与热带医学学院 微生物学系,广东广州510515

出　　处：《南方医科大学学报》2006年第4期469-471,共3页Journal of Southern Medical University

基　　金：广州市科技攻关计划重点项目(2004Z2-E0214);广东省自然科学基金(32833)~~

摘　　要：目的构建登革2型病毒NGC株基因组全序列的亚cDNA克隆,为进一步构建全长感染性cDNA克隆奠定基础。方法根据登革2型病毒NGC株基因组全序列中的酶切位点设计2对引物,从病毒感染的乳鼠脑中提取RNA,采用长链RT-PCR技术扩增出覆盖病毒基因组全长的2个cDNA片段,并克隆至pCR-XL-TOPO载体后测序。结果经酶切鉴定和序列测定表明,获得的cDNA克隆为登革2型病毒NGC株特异的。结论已成功构建出登革2型病毒NGC株基因组的2个cDNA亚克隆。Objective To construct the eDNA subclones spanning the entire genome of dengue 2 virus NGC strain for further construction of full-length infectious viral eDNA clone. Methods Two pairs of primers were designed according to the restriction endonuelease sites in the viral genome of dengue 2 virus NGC strain. After viral RNA extraction from the brain of infected new-born mice, two parts of full-length viral eDNA were amplified by long RT-PCR and cloned into the vector pCR-XL-TOPO. The partial sequence of the recombinant plasmid was determined. Results and Conclusion Sequence analysis and digestion with restriction enzymes demonstrated that the two eDNA subelones were specific for dengue 2 virus NGC strain, suggesting the successful construction of the two eDNA subelones of dengue 2 virus NGC strain.

关 键 词：登革2型病毒 RT-PCR CDNA克隆 

分 类 号：R373.33[医药卫生—病原生物学]