Construction of pGCGi, an expression vector carries intron containing GUS and analysis using micro-bombardment and agroinjection

Transient gene expression is fast, easy and not influenced by positional effects that potentially affect on gene expression levels in stable gene transformation. Transient expression can be applicable using agroinfilteration, biolistic and viral vectors. Agrobacterium mediated transient expression h...

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Main Authors: Mohammad Reza Zamani, Farhad Shokouhifar, Mostafa Motallebi
Format: Article
Language:fas
Published: University of Isfahan 2014-03-01
Series:Zīst/shināsī-i Giyāhī-i Īrān
Subjects:
Online Access:http://ijpb.ui.ac.ir/article_18918_f7fe6ffeca4e21b6ebe504ddec3998b2.pdf
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spelling doaj-c29f127d2c314d9cbcaf80ebbe81f0d52020-11-24T21:28:14ZfasUniversity of IsfahanZīst/shināsī-i Giyāhī-i Īrān2008-82642322-22042014-03-016199711018918Construction of pGCGi, an expression vector carries intron containing GUS and analysis using micro-bombardment and agroinjectionMohammad Reza ZamaniFarhad ShokouhifarMostafa MotallebiTransient gene expression is fast, easy and not influenced by positional effects that potentially affect on gene expression levels in stable gene transformation. Transient expression can be applicable using agroinfilteration, biolistic and viral vectors. Agrobacterium mediated transient expression have been shown as an efficient and versatile method for analyzing transgene expression, gene scilencing, host-pathogen interactions, protein-protein interaction, and cis-element/transfactor interaction. A control vector consists of an interon containing reporter gene under control of a common promoter is often required for Cis-acting elemen analysis to standardize the variation. This study was carried out to construct a control vector based on two parental binary vectors, pGPTV and pCAMBIA3301. The constructed vector, pGCGi had an interon containing GUS reporter gene under control of the full sequence of CaMV 35S promoter. The GUS staining results revealed that the GUS intron containg gene cannot produce an active B- glucronidase enzyme in agrobacterial cells. The functional gene expression analysis of the new vector was done using agroinjection and prticle delivery system in tobacco and onion eoidermal cells respectively. The histochemical GUS staining results showed pGCGi could express the reporter gene in plant cells and culd be used as control vector in transient expression experiments.http://ijpb.ui.ac.ir/article_18918_f7fe6ffeca4e21b6ebe504ddec3998b2.pdfBinary vectorsMicro-bombardmentTransient gene expressionAgroinfiltrationMicrobombardmentBiolistic gene delivery systemGUS staining
collection DOAJ
language fas
format Article
sources DOAJ
author Mohammad Reza Zamani
Farhad Shokouhifar
Mostafa Motallebi
spellingShingle Mohammad Reza Zamani
Farhad Shokouhifar
Mostafa Motallebi
Construction of pGCGi, an expression vector carries intron containing GUS and analysis using micro-bombardment and agroinjection
Zīst/shināsī-i Giyāhī-i Īrān
Binary vectors
Micro-bombardment
Transient gene expression
Agroinfiltration
Micro
bombardment
Biolistic gene delivery system
GUS staining
author_facet Mohammad Reza Zamani
Farhad Shokouhifar
Mostafa Motallebi
author_sort Mohammad Reza Zamani
title Construction of pGCGi, an expression vector carries intron containing GUS and analysis using micro-bombardment and agroinjection
title_short Construction of pGCGi, an expression vector carries intron containing GUS and analysis using micro-bombardment and agroinjection
title_full Construction of pGCGi, an expression vector carries intron containing GUS and analysis using micro-bombardment and agroinjection
title_fullStr Construction of pGCGi, an expression vector carries intron containing GUS and analysis using micro-bombardment and agroinjection
title_full_unstemmed Construction of pGCGi, an expression vector carries intron containing GUS and analysis using micro-bombardment and agroinjection
title_sort construction of pgcgi, an expression vector carries intron containing gus and analysis using micro-bombardment and agroinjection
publisher University of Isfahan
series Zīst/shināsī-i Giyāhī-i Īrān
issn 2008-8264
2322-2204
publishDate 2014-03-01
description Transient gene expression is fast, easy and not influenced by positional effects that potentially affect on gene expression levels in stable gene transformation. Transient expression can be applicable using agroinfilteration, biolistic and viral vectors. Agrobacterium mediated transient expression have been shown as an efficient and versatile method for analyzing transgene expression, gene scilencing, host-pathogen interactions, protein-protein interaction, and cis-element/transfactor interaction. A control vector consists of an interon containing reporter gene under control of a common promoter is often required for Cis-acting elemen analysis to standardize the variation. This study was carried out to construct a control vector based on two parental binary vectors, pGPTV and pCAMBIA3301. The constructed vector, pGCGi had an interon containing GUS reporter gene under control of the full sequence of CaMV 35S promoter. The GUS staining results revealed that the GUS intron containg gene cannot produce an active B- glucronidase enzyme in agrobacterial cells. The functional gene expression analysis of the new vector was done using agroinjection and prticle delivery system in tobacco and onion eoidermal cells respectively. The histochemical GUS staining results showed pGCGi could express the reporter gene in plant cells and culd be used as control vector in transient expression experiments.
topic Binary vectors
Micro-bombardment
Transient gene expression
Agroinfiltration
Micro
bombardment
Biolistic gene delivery system
GUS staining
url http://ijpb.ui.ac.ir/article_18918_f7fe6ffeca4e21b6ebe504ddec3998b2.pdf
work_keys_str_mv AT mohammadrezazamani constructionofpgcgianexpressionvectorcarriesintroncontaininggusandanalysisusingmicrobombardmentandagroinjection
AT farhadshokouhifar constructionofpgcgianexpressionvectorcarriesintroncontaininggusandanalysisusingmicrobombardmentandagroinjection
AT mostafamotallebi constructionofpgcgianexpressionvectorcarriesintroncontaininggusandanalysisusingmicrobombardmentandagroinjection
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