Gong Shu-ji, Cao Hong, Zhao Wei, Zhang Wen-bing, Zhou Hao, Chen Li-dan
Department of Microbiology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2006 Apr;26(4):469-71.
To construct the cDNA subclones spanning the entire genome of dengue 2 virus NGC strain for further construction of full-length infectious viral cDNA clone.
Two pairs of primers were designed according to the restriction endonuclease 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 cDNA were amplified by long RT-PCR and cloned into the vector pCR-XL-TOPO. The partial sequence of the recombinant plasmid was determined.
Sequence analysis and digestion with restriction enzymes demonstrated that the two cDNA subclones were specific for dengue 2 virus NGC strain, suggesting the successful construction of the two cDNA subclones of dengue 2 virus NGC strain.
构建覆盖登革2型病毒NGC株全基因组的cDNA亚克隆，以便进一步构建全长感染性病毒cDNA克隆。
根据登革2型病毒NGC株病毒基因组中的限制性内切酶位点设计两对引物。从感染新生小鼠的脑组织中提取病毒RNA后，通过长链RT-PCR扩增两部分全长病毒cDNA，并克隆到载体pCR-XL-TOPO中。测定重组质粒的部分序列。
序列分析和限制性酶切表明，这两个cDNA亚克隆对登革2型病毒NGC株具有特异性，提示成功构建了登革2型病毒NGC株的两个cDNA亚克隆。
