Cargando…

Construction of eukaryotic expression vector of TSLC1 gene

INTRODUCTION: To construct a eukaryotic expression vector of the tumour suppressor in lung cancer 1 (TSLC1) gene, so as to explore the mechanisms of tumour suppression of the gene theoretically. MATERIAL AND METHODS: The open reading frame (ORF) of TSLC1 gene was amplified with RT-PCR from normal hu...

Descripción completa

Detalles Bibliográficos
Autores principales: Liang, Qi-Lian, Wang, Bi-Rong, Li, Zhou-Yu, Chen, Guo-Qiang, Zhou, Yuan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Termedia Publishing House 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3258782/
https://www.ncbi.nlm.nih.gov/pubmed/22291791
http://dx.doi.org/10.5114/aoms.2011.24124
_version_ 1782221315142320128
author Liang, Qi-Lian
Wang, Bi-Rong
Li, Zhou-Yu
Chen, Guo-Qiang
Zhou, Yuan
author_facet Liang, Qi-Lian
Wang, Bi-Rong
Li, Zhou-Yu
Chen, Guo-Qiang
Zhou, Yuan
author_sort Liang, Qi-Lian
collection PubMed
description INTRODUCTION: To construct a eukaryotic expression vector of the tumour suppressor in lung cancer 1 (TSLC1) gene, so as to explore the mechanisms of tumour suppression of the gene theoretically. MATERIAL AND METHODS: The open reading frame (ORF) of TSLC1 gene was amplified with RT-PCR from normal human foreskin acrobystia, and cloned to pMD19-T simple vector (TA Clone method). The resultant plasmid was transformed into Escherichia coli JM109 for amplification. The TA Clone recombinant was digested by double restriction enzyme (Bgl II/EcoR I) and analysed with agarose gel electrophoresis. The positive one was sequenced. The inserted DNA fragment was recovered, and then it was mounted into the eukaryotic expression vector pIRES2-EGFP, transformed into E. coli JM109 for amplification. A positive recombinant plasmid named pIRES2-EGFP-TSLC1 was confirmed by Bgl II/EcoR I double-enzyme digestion analysis. RESULTS: RT-PCR amplified the ORF of the TSLC1 gene. It was approximately 1400 base pairs. The obtained DNA was confirmed a high degree of homology with the sequence of TSLC1 cDNA sequence (AY358334) stored at GenBank. CONCLUSIONS: Construction of a TSLC1 eukaryotic expression vector was successful, and it has established a solid foundation for further study.
format Online
Article
Text
id pubmed-3258782
institution National Center for Biotechnology Information
language English
publishDate 2011
publisher Termedia Publishing House
record_format MEDLINE/PubMed
spelling pubmed-32587822012-01-30 Construction of eukaryotic expression vector of TSLC1 gene Liang, Qi-Lian Wang, Bi-Rong Li, Zhou-Yu Chen, Guo-Qiang Zhou, Yuan Arch Med Sci Basic Research INTRODUCTION: To construct a eukaryotic expression vector of the tumour suppressor in lung cancer 1 (TSLC1) gene, so as to explore the mechanisms of tumour suppression of the gene theoretically. MATERIAL AND METHODS: The open reading frame (ORF) of TSLC1 gene was amplified with RT-PCR from normal human foreskin acrobystia, and cloned to pMD19-T simple vector (TA Clone method). The resultant plasmid was transformed into Escherichia coli JM109 for amplification. The TA Clone recombinant was digested by double restriction enzyme (Bgl II/EcoR I) and analysed with agarose gel electrophoresis. The positive one was sequenced. The inserted DNA fragment was recovered, and then it was mounted into the eukaryotic expression vector pIRES2-EGFP, transformed into E. coli JM109 for amplification. A positive recombinant plasmid named pIRES2-EGFP-TSLC1 was confirmed by Bgl II/EcoR I double-enzyme digestion analysis. RESULTS: RT-PCR amplified the ORF of the TSLC1 gene. It was approximately 1400 base pairs. The obtained DNA was confirmed a high degree of homology with the sequence of TSLC1 cDNA sequence (AY358334) stored at GenBank. CONCLUSIONS: Construction of a TSLC1 eukaryotic expression vector was successful, and it has established a solid foundation for further study. Termedia Publishing House 2011-08 2011-09-02 /pmc/articles/PMC3258782/ /pubmed/22291791 http://dx.doi.org/10.5114/aoms.2011.24124 Text en Copyright © 2011 Termedia & Banach http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-Noncommercial 3.0 Unported License, permitting all non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Basic Research
Liang, Qi-Lian
Wang, Bi-Rong
Li, Zhou-Yu
Chen, Guo-Qiang
Zhou, Yuan
Construction of eukaryotic expression vector of TSLC1 gene
title Construction of eukaryotic expression vector of TSLC1 gene
title_full Construction of eukaryotic expression vector of TSLC1 gene
title_fullStr Construction of eukaryotic expression vector of TSLC1 gene
title_full_unstemmed Construction of eukaryotic expression vector of TSLC1 gene
title_short Construction of eukaryotic expression vector of TSLC1 gene
title_sort construction of eukaryotic expression vector of tslc1 gene
topic Basic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3258782/
https://www.ncbi.nlm.nih.gov/pubmed/22291791
http://dx.doi.org/10.5114/aoms.2011.24124
work_keys_str_mv AT liangqilian constructionofeukaryoticexpressionvectoroftslc1gene
AT wangbirong constructionofeukaryoticexpressionvectoroftslc1gene
AT lizhouyu constructionofeukaryoticexpressionvectoroftslc1gene
AT chenguoqiang constructionofeukaryoticexpressionvectoroftslc1gene
AT zhouyuan constructionofeukaryoticexpressionvectoroftslc1gene