SARS-CoV 5’-UTR相应cDNA作启动子时转录起始位点研究

张建军; 黄爱龙; 史小玲; 张晓锋

doi:10.13406/j.cnki.cyxb.2005.06.019

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SARS-CoV 5’-UTR相应cDNA作启动子时转录起始位点研究

更新时间：2024-01-17

- SARS-CoV 5’-UTR相应cDNA作启动子时转录起始位点研究
- Localization of the initiation site of the transcription driven by the cDNA corresponding to SARS - CoV 5’ - UTR
- 重庆医科大学学报 2005年第6期页码：849-851
- 作者机构：
  
  1. 感染性疾病分子生物学教育部重点实验室、重庆医科大学病毒性肝炎研究所
  2. 泸州医学院附属医院感染病科
  3. 泸州医学院附属医院感染病科重庆 400016 泸州医学院附属医院感染病科
  4. ,泸州,646000
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2005.06.019
  中图分类号： R373
- 纸质出版日期：2005，
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[1]张建军,黄爱龙,史小玲,张晓锋.SARS-CoV 5’-UTR相应cDNA作启动子时转录起始位点研究[J].重庆医科大学学报,2005(06):849-851.

Zhang Jianjun, et al. Localization of the initiation site of the transcription driven by the cDNA corresponding to SARS - CoV 5’ - UTR[J]. Journal of Chongqing Medical University, 2005,(6):849-851.
[1]张建军,黄爱龙,史小玲,张晓锋.SARS-CoV 5’-UTR相应cDNA作启动子时转录起始位点研究[J].重庆医科大学学报,2005(06):849-851. DOI： 10.13406/j.cnki.cyxb.2005.06.019.

Zhang Jianjun, et al. Localization of the initiation site of the transcription driven by the cDNA corresponding to SARS - CoV 5’ - UTR[J]. Journal of Chongqing Medical University, 2005,(6):849-851. DOI： 10.13406/j.cnki.cyxb.2005.06.019.

摘要

目的:分析具有启动子功能的SARS-CoV基因组5’-UTR cDNA在转录出mRNA时

转录起始的位点

从而分析序列功能。方法将SARS-CoV 5’-UTR驱动下报告基因的真核表达质粒pGL3-5’-UTR

转染HepG2细胞

然后提取总 RNA

采用5’-RACE

扩增出相应序列

通过比照

查找转录其始的位点。结果通过逆转录和二轮PCR

得到一个约440bp 的产物

经A-T克隆后测序

转录体起始位点在SARS-CoV 5’-UTR的第56位核苷酸

其下游是保守的保守的转录调控序列（TRS）。结论SARS-CoV 5’-UTR在调控基因转录时

第56位核苷酸及其下游的TRS具有重要作用。

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.

关键词

SARS-CoV5’-非编码区启动子起始位点

Keywords

SARS-CoV5’ -UTRpromoterInitiation site

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