根癌农杆菌介导深红虫草菌株C033转化体系的建立

生物技术通报 ›› 2013, Vol. 0 ›› Issue (5): 149-154.

根癌农杆菌介导深红虫草菌株C033转化体系的建立

郭维^{1, 2}, 王磊¹, 吴宏清^{1, 2}, 白玲², 章卫民¹

(1.广东省微生物研究所广东省菌种保藏与应用重点实验室广东省微生物应用新技术公共实验室广东省华南应用微生物重点实验室-省部共建国家重点实验室培育基地，广州 510070；2.江西农业大学理学院，南昌 330045)

收稿日期:2012-12-21 修回日期:2013-05-24 出版日期:2013-05-24 发布日期:2013-05-24
作者简介:郭维，女，硕士研究生，研究方向：微生物分子生物学；E-mail：731564955@qq.com
基金资助:
国家自然科学基金项目(30870450)

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 Published:2013-05-24 Online: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

摘要/Abstract

摘要： 利用根癌农杆菌EHA105介导，建立了深红虫草(Cordyceps cardinalis)菌株C033的遗传转化体系。遗传霉素(G418)的抗性筛选，gus基因活性检测和PCR分析结果表明，nptⅡ抗性筛选标记基因已经整合到转化子基因组DNA中，并能够稳定遗传；同时对影响该菌株的转化因素包括孢子浓度、农杆菌OD值、乙酰丁香酮(AS)的浓度及共培养时间进行分析，在优化条件下的转化效率为100个转化子/10⁵个孢子。

关键词: 深红虫草, 根癌农杆菌, 转化体系, nptⅡ基因, gus基因

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

郭维, 王磊, 吴宏清, 白玲, 章卫民. 根癌农杆菌介导深红虫草菌株C033转化体系的建立[J]. 生物技术通报, 2013, 0(5): 149-154.

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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