Construction and identification of genomic cDNA subclones of dengue 2 virus NGC strain.
- Author:
Shu-ji GONG
1
;
Hong CAO
;
Wei ZHAO
;
Wen-bing ZHANG
;
Hao ZHOU
;
Li-dan CHEN
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Animals, Newborn; Brain; virology; Cloning, Molecular; DNA, Complementary; biosynthesis; genetics; DNA, Viral; biosynthesis; genetics; Dengue; virology; Dengue Virus; classification; genetics; isolation & purification; Genome, Viral; Mice; RNA, Viral; genetics; isolation & purification; Recombination, Genetic; Reverse Transcriptase Polymerase Chain Reaction
- From: Journal of Southern Medical University 2006;26(4):469-471
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct the cDNA subclones spanning the entire genome of dengue 2 virus NGC strain for further construction of full-length infectious viral cDNA clone.
METHODSTwo 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 CONCLUSIONSequence 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.
