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




A Mutagenesis Assay for Reporter Gene Screening Using Partially Degenerate Oligonucleotides of the Tandems NNT and NNC - Download this document for free, or read online. Document in PDF available to download.

BioMed Research International - Volume 2015 2015, Article ID 950873, 7 pages -

Research Article

Department of Molecular Diagnostics and Biopharmaceutics, College of Pharmaceutical Science, Soochow University, Suzhou 215123, China

Department of Human Anatomy, University of South China, Hengyang 421001, China

Received 7 January 2015; Accepted 2 March 2015

Academic Editor: Xian-Wei Liu

Copyright © 2015 Huifen Xu et al. 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.

Abstract

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.





Author: Huifen Xu, Cuilan Zhou, Andy K. Zhang, Wen Li, Jia Zhang, and Kai Li

Source: https://www.hindawi.com/



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