Localization of the initiation site of the transcription driven by the cDNA corresponding to SARS - CoV 5' - UTR
- VernacularTitle:SARS-CoV 5’-UTR相应cDNA作启动子时转录起始位点研究
- Author:
Jianjun ZHANG
- Publication Type:Journal Article
- Keywords:
SARS-CoV, 5' -UTR, Promoter, Initiation site
- From:
Journal of Chongqing Medical University
2003;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objectives To localize the initiation site of the transcript driven by the cDNA corresponding to SARS - CoV 5 ' - UTR when it act as a promoter. Methods The plasmid pGI3 -5' - UTR, in which a reporter gene Luc was driven by the cDNA corresponding to SARS - CoV 5' -UTR, was transfected into HepG2 cells, and then the total RNA was extracted. Subsequently, the extracted total RNA was reverse - transcribed using a specific primer and subjected to two turns of semi - nested PCR. The PCR product was cloned into a T - vector for sequencing. Results A 440bps product was harvested by reverse - transcription and two turns of semi - nested PCR, and the 56th nucleotide of SARS - CoV 5 ' - UTR was found being the initiation site, followed downstream with a highly conserved transcription - regulating sequence (TRS). Conclusion The 56th nucleotide and its downstream TRS of SARS - CoV 5 ' - UTR is key roles in regulating gene transcription.
