阿扎霉素F产生菌链霉菌211726基因转移系统的建立

doi:10.13560/j.cnki.biotech.bull.1985.2016.04.026

生物技术通报 ›› 2016, Vol. 32 ›› Issue (4): 198-202.doi: 10.13560/j.cnki.biotech.bull.1985.2016.04.026

阿扎霉素F产生菌链霉菌211726基因转移系统的建立

马艳玲¹,刘富来¹,张敏¹,孙宇辉²,洪葵²,

1. 佛山科学技术学院食品与园艺学院，佛山 528231;
2. 武汉大学药学院，武汉 430071

收稿日期:2015-07-23 出版日期:2016-04-25 发布日期:2016-04-26
作者简介:马艳玲，女，博士，研究方向：食品生物技术；E-mail：mayanling100@163.com
基金资助:
国家自然科学基金项目（31270120），佛山市科技计划项目（2014GA000425，2014AG10007）

The Construction of the Gene Transfer System of Strain Streptomyces sp. 211726 Producing Azalomycin F

MA Yan-ling¹, LIU Fu-lai¹, ZHANG Min¹, SUN Yu-hui², HONG Kui²

1. College of Food and Horticultural Sciences，Foshan University，Foshan 528231;
2. School of Pharmaceutical Sciences，Wuhan University，Wuhan 430071

Received:2015-07-23 Published:2016-04-25 Online:2016-04-26

摘要/Abstract

摘要：

旨在建立阿扎霉素F产生菌链霉菌211726的基因转移系统，以便基因敲除和外源基因表达等遗传操作。以整合型质粒pSET152和pIB139为出发质粒，通过接合转移构建了阿扎霉素F产生菌链霉菌211726的基因转移系统。结果显示25μg/mL阿泊拉霉素可有效筛选接合子。经PCR验证，质粒成功整合到菌株链霉菌211726基因组中，接合子经多次传代后，导入的质粒 pSET152和pIB139仍稳定整合于接合子基因组上。

关键词: 阿扎霉素F, 链霉菌211726, 接合转移, 遗传稳定性

Abstract:

The objective of this study is to establish the gene transfer system of strain Streptomyces sp. 211726 producing azalomycin F，which can be used for genetic manipulations such as gene knock-out and expression of foreign genes. Intergeneric genetic transfer system of Streptomyces sp. 211726 producing azalomycin F was constructed by conjugating integrative plasmid pSET152 with pIB139. Results showed that 25 mg/mL apramycin may be used to efficiently screen conjugants. PCR verification revealed that exogenous plasmid was successfully integrated in the chromosomal DNA of Streptomyces sp. 211726. The continuous passage culture experiment demonstrated that transformed pSET152 and pIB139 of conjugants were stably inherited.

Key words: azalomycin F, Streptomyces sp. 211726, conjugation, genetic stability

马艳玲,刘富来,张敏,孙宇辉,洪葵. 阿扎霉素F产生菌链霉菌211726基因转移系统的建立[J]. 生物技术通报, 2016, 32(4): 198-202.

MA Yan-ling, LIU Fu-lai, ZHANG Min, SUN Yu-hui, HONG Kui. The Construction of the Gene Transfer System of Strain Streptomyces sp. 211726 Producing Azalomycin F[J]. Biotechnology Bulletin, 2016, 32(4): 198-202.

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阿扎霉素F产生菌链霉菌211726基因转移系统的建立

The Construction of the Gene Transfer System of Strain Streptomyces sp. 211726 Producing Azalomycin F

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