电穿孔介导马尔尼菲青霉转化体系的建立和优化

中国真菌学杂志　 2014, Vol. 9 　Issue (1): 12-15.

电穿孔介导马尔尼菲青霉转化体系的建立和优化

陈阳霞¹, 顾桉菁², 梁宇恒¹, 马团¹, 席丽艳¹

1. 中山大学孙逸仙纪念医院, 广州 510120;
2. 贵阳医学院基础医学院, 贵阳 550004

收稿日期:2013-12-21 出版日期:2014-02-28 发布日期:2014-02-28
通讯作者: 席丽艳，E-mail：xiliyan@mail.sysu.edu.cn E-mail:xiliyan@mail.sysu.edu.cn
作者简介:陈阳霞，女（苗族），硕士研究生在读.E-mail：chenyangxiasmu@126.com
基金资助:
国家自然科学基金（81171545）

Electroporation-mediate transformation of Penicillium marneffei and optimization of the transforming conditions

CHEN Yang-xia¹, GU An-jing², LIANG Yu-heng¹, MA Tuan¹, XI Li-yan¹

1. Department of Dermatology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, China;
2. College of Basic Medicine, Guiyang Medical University, Guiyang 550004, China

Received:2013-12-21 Online:2014-02-28 Published:2014-02-28

摘要/Abstract

摘要： 目的建立和优化马尔尼菲青霉电穿孔转化体系，为其基因功能研究提供良好平台。方法直接使用马尔尼菲青霉缺陷株SPM4（pyrG^-，niaD^-）萌发孢子进行电穿孔转化，并通过改变孢子龄、孢子萌发时间、质粒浓度、电场强度等影响因素对体系进行优化。结果适合SPM4电穿孔转化条件为：孢子龄为6 d，孢子萌发时间为4 h，电场强度为5 kV/cm。上述条件下分别使用1 μg环状或线性化质粒DNA转化SPM4，平均可得到21个和13个转化子。结论马尔尼菲青霉电穿孔转化效率高，重复性好，针对SPM4萌发孢子，环状质粒较线性化质粒电穿孔转化效率高。

关键词: 马尔尼菲青霉, 萌发孢子, 电穿孔

Abstract: Objective To establish and optimize the electroporation-mediated transformation system for Penicillium marneffei.It would provide a good platform for the gene function research of P.marneffei.Method The selectable marker pyrG gene was transformed into the germinated spores of Penicillium marneffei strain SPM4 (pyrG^-,niaD^-) by electroporation method. It aimed at finding out the optimal protocol by changing some important effecting factors including spore stadium,germination time,electric field intensity, and so on.Results The appropriate electroporation conditions for SPM4 were as follows: spore stadium 6 d, germination time 4 h and electric field intensity 5 kV/cm. Under these conditions, 21 and 13 transformants were obtained using 1 μg circular and linearized plasmid DNA, separately.Conclusion Electroporation-mediate transformation of Penicillium marneffei can get high transformation efficiency and repeatability. It is implied that circular plasmid got higher transformation frequency than the linearized by electroporation for the germinated spores of SPM4.

Key words: Penicillium marneffei, germinated spores, electroporation

中图分类号:

R379.9

陈阳霞, 顾桉菁, 梁宇恒, 马团, 席丽艳. 电穿孔介导马尔尼菲青霉转化体系的建立和优化[J]. 中国真菌学杂志, 2014, 9(1): 12-15.

CHEN Yang-xia, GU An-jing, LIANG Yu-heng, MA Tuan, XI Li-yan. Electroporation-mediate transformation of Penicillium marneffei and optimization of the transforming conditions[J]. Chinese Journal of Mycology, 2014, 9(1): 12-15.

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