Construction and analysis of activity of an HIV-1/bovine immunodeficiency virus chimeric clone cDNA.
- Author:
Yi-shu YANG
1
;
Guo-min CHEN
;
Wen-ping DONG
;
Qi-min CHEN
;
Yun-qi GENG
;
Yi ZENG
Author Information
- Publication Type:Journal Article
- MeSH: AIDS Vaccines; Animals; Cattle; Cloning, Molecular; DNA, Complementary; genetics; DNA, Viral; genetics; Genes, gag; genetics; Genes, pol; genetics; Genes, tat; genetics; HIV-1; genetics; Humans; Immunodeficiency Virus, Bovine; genetics; Recombinant Fusion Proteins; genetics; metabolism; Transcription, Genetic; Transcriptional Activation; Transfection; Virus Replication
- From: Chinese Journal of Experimental and Clinical Virology 2003;17(2):143-145
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEChimeric human/bovine immunodeficiency virus (HBIV) cDNA was constructed by replacing HIV tat and LTR with bovine immunodeficiency virus (BIV) tat and LTR to study the activity of BIV tat and LTR in the chimerae.
METHODSThe target fragments of BIV tat, LTR and HIV gag, pol, env were respectively amplified by using PCR and sequentially inserted into pBluescript SK(+) vector. The chimeric clone was transfected into human MT4 cells. The transcript and gene expression of the HBIV chimeric virus were detected by using RT-PCR and a reverse transcriptase assay, respectively.
RESULTSBIV tat mRNA and HIV gag mRNA were detected. The reverse transcriptase activity of the chimeric virus was analyzed in the fluctuation curve.
CONCLUSIONSIn chimeric HBIV cDNA transfected MT?4 cells, BIV tat and HIV gag were transcripted. The reverse transcriptase of the chimeric virus had biological activity. These data suggest that in MT4 cells, BIV LTR had promoter activity and BIV tat had the function of transactivation in the chimeric virus. The study of the chimeric virus with infectivity is in progress.
