A Mutagenesis Assay for Reporter Gene Screening Using Partially Degenerate Oligonucleotides of the Tandems NNT and NNC

Not all proteins are tolerable to mutations. Whether a specific protein can be a mutable target is of importance in the biotechnology and pharmaceutical industry. This study reported a novel mutagenesis assay using tandem NNT and NNC oligonucleotides to test the mutability of a candidate gene. These...

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Detalles Bibliográficos
Autores principales: Xu, Huifen, Zhou, Cuilan, Zhang, Andy K., Li, Wen, Zhang, Jia, Li, Kai
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2015
Materias:
Research Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4488517/
https://www.ncbi.nlm.nih.gov/pubmed/26167508
http://dx.doi.org/10.1155/2015/950873
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author Xu, Huifen
Zhou, Cuilan
Zhang, Andy K.
Li, Wen
Zhang, Jia
Li, Kai
author_facet Xu, Huifen
Zhou, Cuilan
Zhang, Andy K.
Li, Wen
Zhang, Jia
Li, Kai
author_sort Xu, Huifen
collection PubMed
description Not all proteins are tolerable to mutations. Whether a specific protein can be a mutable target is of importance in the biotechnology and pharmaceutical industry. This study reported a novel mutagenesis assay using tandem NNT and NNC oligonucleotides to test the mutability of a candidate gene. These two tandem oligonucleotides avoid the risk of forming nonsense mutations and render flexibility of truncating or expanding the insertion size. As a reporter gene, ZeoR (zeocin resistance gene) was confirmed to have a high tolerance for mutagenesis by this new assay.
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spelling pubmed-44885172015-07-12 A Mutagenesis Assay for Reporter Gene Screening Using Partially Degenerate Oligonucleotides of the Tandems NNT and NNC Xu, Huifen Zhou, Cuilan Zhang, Andy K. Li, Wen Zhang, Jia Li, Kai Biomed Res Int Research Article Not all proteins are tolerable to mutations. Whether a specific protein can be a mutable target is of importance in the biotechnology and pharmaceutical industry. This study reported a novel mutagenesis assay using tandem NNT and NNC oligonucleotides to test the mutability of a candidate gene. These two tandem oligonucleotides avoid the risk of forming nonsense mutations and render flexibility of truncating or expanding the insertion size. As a reporter gene, ZeoR (zeocin resistance gene) was confirmed to have a high tolerance for mutagenesis by this new assay. Hindawi Publishing Corporation 2015 2015-06-18 /pmc/articles/PMC4488517/ /pubmed/26167508 http://dx.doi.org/10.1155/2015/950873 Text en Copyright © 2015 Huifen Xu et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Xu, Huifen
Zhou, Cuilan
Zhang, Andy K.
Li, Wen
Zhang, Jia
Li, Kai
A Mutagenesis Assay for Reporter Gene Screening Using Partially Degenerate Oligonucleotides of the Tandems NNT and NNC
title A Mutagenesis Assay for Reporter Gene Screening Using Partially Degenerate Oligonucleotides of the Tandems NNT and NNC
title_full A Mutagenesis Assay for Reporter Gene Screening Using Partially Degenerate Oligonucleotides of the Tandems NNT and NNC
title_fullStr A Mutagenesis Assay for Reporter Gene Screening Using Partially Degenerate Oligonucleotides of the Tandems NNT and NNC
title_full_unstemmed A Mutagenesis Assay for Reporter Gene Screening Using Partially Degenerate Oligonucleotides of the Tandems NNT and NNC
title_short A Mutagenesis Assay for Reporter Gene Screening Using Partially Degenerate Oligonucleotides of the Tandems NNT and NNC
title_sort mutagenesis assay for reporter gene screening using partially degenerate oligonucleotides of the tandems nnt and nnc
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4488517/
https://www.ncbi.nlm.nih.gov/pubmed/26167508
http://dx.doi.org/10.1155/2015/950873
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