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中国农学通报

中国农学通报 ›› 2013, Vol. 29 ›› Issue (9): 155-158.doi: 10.11924/j.issn.1000-6850.2012-3471

• 生物技术科学 • 上一篇    下一篇

不吸水链霉菌公主岭变种 769原生质体制备条件初探

张振鲁 杜茜 周幸 董宝池 李启云 王金刚   

  • 收稿日期:2012-10-27 修回日期:2012-11-23 出版日期:2013-03-25 发布日期:2013-03-25
  • 基金资助:
    中青年科技领军人才及优秀创新团队项目“吉林省农业微生物研究与利用研究创新团队”(20121812)；吉林省农业微生物重点实验室平台建设项目(20122105)；吉林省农科院植保所青年基金项目(zbs2010-01)。

A Preliminary Study on Protoplast Preparation Conditions of Streptomyces gongzhulingensis 769

  • Received:2012-10-27 Revised:2012-11-23 Online:2013-03-25 Published:2013-03-25

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摘要： 为获得较高的不吸水链霉菌公主岭变种769（简称链霉菌769）原生质体产率，采用酶解法制备原生质体，通过单因素试验法探索链霉菌769原生质体制备的最佳条件，包括溶菌酶浓度、酶解时间、酶解温度和渗透压稳定剂。试验结果表明，链霉菌769孢子在溶菌酶浓度为3.5 mg/mL，酶解温度为36℃，酶解时间3 h，以P缓冲液为渗透压稳定剂的条件下原生质体形成效果最好，原生质体再生率达到62.5%。链霉菌769原生质体的获得为以原生质体为基础的基因转移系统的建立奠定了基础。

关键词: ELES模型, ELES模型

Abstract: In order to obtain high yields of the protoplasts of Streptomyces gongzhulingensis 769, this paper described an enzymatic method for that. In this study, spores of S. gongzhulingensis 769 were treated with bacteriolytic enzyme. The optimal conditions, including the concentration of lysozyme, enzymolysis time, enzymolysis temperature and the osmotic stabilizer, were studied via single-factor method. The result indicated that when the spores were treated with bacteriolytic enzyme of 3.5 mg/mL, at 36℃for 3 hours in osmotic stabilizer of P buffer, the regeneration rate of the protoplasts can reach 62.5%. And the protoplasts preparation makes important foundation for the establishing of the protoplast-mediated genetic transformation system of S. gongzhulingensis 769.