红曲霉不同转化方法比较

遗传 ›› 2006, Vol. 28 ›› Issue (4): 479-485.

红曲霉不同转化方法比较

周礼红¹;诸葛健²

1. 贵州大学生命科学院，贵阳花溪 550025；2. 江南大学生物工程学院工业微生物研究中心，工业生物技术教育部重点实验室，无锡 214036

收稿日期:2005-04-21 修回日期:2005-06-12 出版日期:2006-04-10 发布日期:2006-04-10
通讯作者: 诸葛健

Comparison of Different Transformation Methods for Monascus sp.

ZHOU Li-Hong¹，ZHUGE Jian²，WANG Zheng-Xiang²

1. Department of biotechnology, Guizhou University, Huaxi Guiyang 550025, China；2. Research Center of Industrial Microbiology, School of Biotechnology,The Key Laboratory of Industrial Biotechnology, Ministry of Education, Southern Yangtze University, Wuxi 214036, China

Received:2005-04-21 Revised:2005-06-12 Online:2006-04-10 Published:2006-04-10
Contact: ZHUGE Jian

摘要/Abstract

摘要：

为了研究红曲霉聚酮体途径，考察和比较了4种不同的转化方法以建立有效的红曲霉遗传转化系统。以潮霉素作为抗性筛选标记，pBC-Hygro作为转化载体，用基于原生质体的传统转化和电击转化、基于萌发孢子的电击转化以及REMI技术转化红曲霉。发现基于萌发孢子的电击转化由于转化率极低而不适于红曲霉转化。基于原生质体的传统转化和电击转化尽管每微克DNA分别能获得135个转化子和125个转化子，但因转化子稳定性差也适合红曲霉转化的转化。应用REMI技术，转化率提高约20倍，每微克DNA 2500个转化子，70%～75%的转化子的稳定，非常适合于红曲霉的转化。

关键词: 红曲霉, 遗传转化, 电击转化, REMI

Abstract: In order to facilitate the producer of polyketide pathway, four different transformation methods were tested and compared in an attempt to develop the genetic transformation system of Monascus sp.. Using vector pBC-Hygro, the fungus was transformed to be hygromycin B- resistant, by conventional transformation as well as electroporation based on protoplast, electroporation based on germinated conidia, and restriction enzyme-mediated integration (REMI). Electroporation based on germinated conidia was found to be inappropriate for transforming Monascus sp. due to a low transformation frequency. The conventional transformation and electroporation technique based on protoplasts were thought not to be fit for transforming Monascus sp., due to a low stability of transformants though they yielded up to 135 transformants and 125 transformants per microgrammol/Lol/Le DNA, respectively. Transformant number was increased by 20-fold by REMI (2500 transformants per microgrammol/Lol/Le DNA) and 70%～75％ of them were stable. REMI technique would be very beneficial to the establishment of the

中图分类号:

Q933

周礼红，诸葛健. 红曲霉不同转化方法比较[J]. 遗传, 2006, 28(4): 479-485.

ZHOU Li-Hong, ZHUGE Jian, WANG Zheng-Xiang. Comparison of Different Transformation Methods for Monascus sp.[J]. HEREDITAS, 2006, 28(4): 479-485.

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