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


CONCLUSIONS
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.
Gong
Shu-ji
SJ
Department of Microbiology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China. gongshuji2004@yahoo.com.cn
Cao
Hong
H
Zhao
Wei
W
Zhang
Wen-bing
WB
Zhou
Hao
H
Chen
Li-dan
LD
D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000821:virology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics*; D004279:DNA, Viral / Q000096:biosynthesis / Q000235:genetics*; D003715:Dengue / Q000821:virology; D003716:Dengue Virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D016679:Genome, Viral; D051379:Mice; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D011995:Recombination, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018076:DNA, Complementary; D004279:DNA, Viral; D012367:RNA, Viral