Transformation System of Cordyceps cardinalis Strain C033 Mediated by Agrobacterium tumefaciens

Biotechnology Bulletin ›› 2013, Vol. 0 ›› Issue (5): 149-154.

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Transformation System of Cordyceps cardinalis Strain C033 Mediated by Agrobacterium tumefaciens

Guo Wei^{1, 2}, Wang Lei¹, Wu Hongqing^{1, 2}, Bai Ling², Zhang Weimin¹

(1. State Key Laboratory of Applied Microbiology Ministry-Guangdong Province Jointly Breeding Base，South China，Guangdong Open Laboratory of Applied Microbiology，Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application，Guangdong Institute of Microbiology，Guangzhou 510070；2. College of Sciences，Jiangxi Agricultural University，Nanchang 330045)

Received:2012-12-21 Revised:2013-05-24 Online:2013-05-24 Published:2013-05-24
About author: 章卫民，男，研究员，研究方向：微生物活性物质及其相关功能基因；E-mail：wmzhang58@yahoo.com.cn白玲，女，副教授；E-mail：bailing716@yahoo.com.cn

Abstract

Abstract: By using Agrobacterium tumefaciens-mediated transformation (ATMT), the transformation system of Cordyceps cardinalis strain C033 with A. tumefaciens EHA105 was successfully established. The results of geneticin-resistance-selection, gus activity detection and PCR analysis showed that nptⅡin T-DNA was integrated into the transformants and the transformants was genetically stable. Meanwhile, the factors affecting the transformation efficiency such as the concentration of strain C033 spores, the OD value of A. tumefaciens, the concentration of acetoyringone (AS) and the co-culture time were analyzed. The transformation efficiency was about 100 transformants per 10⁵ spores under the optimal condition. The application of ATMT in C. cardinalis will provide a powerful tool for the functional gene analysis of this fungus.

Key words: Cordyceps cardinalis, Agrobacterium tumefaciens, Transformation system, nptⅡ gene, gus gene

Guo Wei, Wang Lei, Wu Hongqing, Bai Ling, Zhang Weimin. Transformation System of Cordyceps cardinalis Strain C033 Mediated by Agrobacterium tumefaciens[J]. Biotechnology Bulletin, 2013, 0(5): 149-154.

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Transformation System of Cordyceps cardinalis Strain C033 Mediated by Agrobacterium tumefaciens

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