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篇目详细内容 |
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【篇名】 |
Improvement of megaprimer method for site-directed mutagenesis and its application to phytase |
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【刊名】 |
Frontiers of Agriculture in China |
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【刊名缩写】 |
Front. Agric. China |
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【ISSN】 |
1673-7334 |
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【EISSN】 |
1673-744X |
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【DOI】 |
10.1007/s11703-009-0010-5 |
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【出版社】 |
Higher Education Press and Springer-Verlag |
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【出版年】 |
2009 |
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【卷期】 |
3
卷1期 |
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【页码】 |
43-46
页,共
4
页 |
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【作者】 |
Haiqiang LU;
Hongwei YU;
Runfang GUO;
Yingmin JIA;
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【关键词】 |
site-directed mutagenesis; phytase; the single-stranded product; PCR |
【摘要】 |
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Site-directed mutagenesis is used extensively for probing gene function. In this paper we describe an improved megaprimer method to the site-directed mutagenesis of phytase from Aspergillus niger, which allowed the mutations to be performed more efficiently in less time than other traditional methods. Three rounds of PCR and two pairs of primers were required in this method, and additionally, the restriction enzyme Dpn I was used for the elimination of template instead of the gel purification in this process. The entire procedure was performed in one tube. Moreover, this method was easier for obtaining large mutant genes than other methods. We successfully carried out the site-directed mutagenesis of phytase by adopting this method. |
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