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

Abstract

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

CLC Number:

R379.9

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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