Journal of Southern Medical University ›› 2006, Vol. 26 ›› Issue (04): 469-174.

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

GONG Shu-ji, CAO Hong, ZHAO Wei, ZHANG Wen-bing, ZHOU Hao, CHEN Li-dan Department of Microbiology, Institute of Virology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China   

  1. 南方医科大学公共卫生与热带医学学院微生物学系; 南方医科大学公共卫生与热带医学学院病毒研究所; 南方医科大学公共卫生与热带医学学院 微生物学系; 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; 广东 广州 510515;
  • Online:2006-04-20 Published:2006-04-20

Abstract: Objective 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. Methods 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. Results and Conclusion 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. 

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