Technique of Overlap Extension by Polymerase Chain Reaction for Splicing Cauliflower Mosaic Virus (CaMV) 35S Promoter and DhPEX11-Like

The promoter plays an important role in the regulation of gene expression. The problem is some of binary vector that absence from promoter at cloning site. The cauliflower mosaic virus (CaMV) 35S promoter is a strong and constitutive promoter that widely used to produce transgenic organisms. In this...

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Main Authors: Silvi Ikawati, Yung Fu Yen
Format: Article
Language:English
Published: University of Brawijaya 2015-06-01
Series:Journal of Experimental Life Science
Subjects:
Online Access:https://jels.ub.ac.id/index.php/jels/article/view/152
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author Silvi Ikawati
Yung Fu Yen
author_facet Silvi Ikawati
Yung Fu Yen
author_sort Silvi Ikawati
collection DOAJ
description The promoter plays an important role in the regulation of gene expression. The problem is some of binary vector that absence from promoter at cloning site. The cauliflower mosaic virus (CaMV) 35S promoter is a strong and constitutive promoter that widely used to produce transgenic organisms. In this experiment the cauliflower mosaic virus (CaMV) 35S promoter was spliced at upstream of DhPEX11-like for driving downstream transgenes DhPEX11-like expression used the technique of Overlap Extension by The Polymerase Chain Reaction. In gene splicing, internal primers are used to amplify some overlapping regions of both genes and then these internal primers are combined with the external primers in PCR process which allows amplification of the entire region. In the experiment, the recombinant PCR successfully spliced the 35S-DhPEX11 gene. This method is simple, rapid and reduced reagents used because it does not need many vector constructions.
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issn 2087-2852
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publishDate 2015-06-01
publisher University of Brawijaya
record_format Article
series Journal of Experimental Life Science
spelling doaj-art-78aea13699a0458083cccd7f193f69792025-08-20T01:55:33ZengUniversity of BrawijayaJournal of Experimental Life Science2087-28522338-16552015-06-01513843https://doi.org/10.21776/ub.jels.2015.005.01.07Technique of Overlap Extension by Polymerase Chain Reaction for Splicing Cauliflower Mosaic Virus (CaMV) 35S Promoter and DhPEX11-LikeSilvi Ikawati0Yung Fu Yen1Universitas Brawijaya, Malang, IndonesiaNational Chiayi University, TaiwanThe promoter plays an important role in the regulation of gene expression. The problem is some of binary vector that absence from promoter at cloning site. The cauliflower mosaic virus (CaMV) 35S promoter is a strong and constitutive promoter that widely used to produce transgenic organisms. In this experiment the cauliflower mosaic virus (CaMV) 35S promoter was spliced at upstream of DhPEX11-like for driving downstream transgenes DhPEX11-like expression used the technique of Overlap Extension by The Polymerase Chain Reaction. In gene splicing, internal primers are used to amplify some overlapping regions of both genes and then these internal primers are combined with the external primers in PCR process which allows amplification of the entire region. In the experiment, the recombinant PCR successfully spliced the 35S-DhPEX11 gene. This method is simple, rapid and reduced reagents used because it does not need many vector constructions.https://jels.ub.ac.id/index.php/jels/article/view/152dhpex11-likegene splicingoligonucleotidepcrpromoter
spellingShingle Silvi Ikawati
Yung Fu Yen
Technique of Overlap Extension by Polymerase Chain Reaction for Splicing Cauliflower Mosaic Virus (CaMV) 35S Promoter and DhPEX11-Like
Journal of Experimental Life Science
dhpex11-like
gene splicing
oligonucleotide
pcr
promoter
title Technique of Overlap Extension by Polymerase Chain Reaction for Splicing Cauliflower Mosaic Virus (CaMV) 35S Promoter and DhPEX11-Like
title_full Technique of Overlap Extension by Polymerase Chain Reaction for Splicing Cauliflower Mosaic Virus (CaMV) 35S Promoter and DhPEX11-Like
title_fullStr Technique of Overlap Extension by Polymerase Chain Reaction for Splicing Cauliflower Mosaic Virus (CaMV) 35S Promoter and DhPEX11-Like
title_full_unstemmed Technique of Overlap Extension by Polymerase Chain Reaction for Splicing Cauliflower Mosaic Virus (CaMV) 35S Promoter and DhPEX11-Like
title_short Technique of Overlap Extension by Polymerase Chain Reaction for Splicing Cauliflower Mosaic Virus (CaMV) 35S Promoter and DhPEX11-Like
title_sort technique of overlap extension by polymerase chain reaction for splicing cauliflower mosaic virus camv 35s promoter and dhpex11 like
topic dhpex11-like
gene splicing
oligonucleotide
pcr
promoter
url https://jels.ub.ac.id/index.php/jels/article/view/152
work_keys_str_mv AT silviikawati techniqueofoverlapextensionbypolymerasechainreactionforsplicingcauliflowermosaicviruscamv35spromoteranddhpex11like
AT yungfuyen techniqueofoverlapextensionbypolymerasechainreactionforsplicingcauliflowermosaicviruscamv35spromoteranddhpex11like