两启动子影响egfp在绿僵菌中的荧光表达

doi:10.3864/j.issn.0578-1752.2015.S.003

Abstract

Abstract: 【Objective】The study aimed to determine the effect of two promoters, PgpdA and RP27, on egfp gene expression. It would be important for construction of engineered Metarhizium anisopliae with a new and high-expression promoter in future. 【Method】Two expression vectors, PAN-r-egfp including promoter RP27 and PAN-p-egfp including promoter PgpdA, were constructed with a hph marker gene and a egfp reporter gene, respectively. CaCl₂-PEG meditated transformation was used to transfer the two vectors into M. anisopliae strainMa9. The transformants were observed fluorescence intensity of hypha and conidia by Laser Scanning Confocal Microscope.【Result】Among of the hph-resistance transformants from PAN-r-egfp and from PAN-p-egfp transformed, 40% and 31% achieved egfp expression with high fluorescence in M. anisopliae, respectively. Both hypha and conidia of transformants from PAN-r-egfp transformation shined strong fluorescence. But only hypha of transformants from PAN-p-egfp transformation was observed fluorescence, its conidia failed to produce fluorescence.【Conclusion】The egfp gene expression was influenced by different promoters and different development stages of M. anisopliae. The promoter RP27 could drive egfp gene expression either in hypha or in conidia of M. anisopliae Ma9. The initiating action of promoter PgpdA may be inhibited at conidiating stage of M. anisopliae Ma9, resulting in failure of egfp expression at the stage.

Key words: egfp, promoter RP27, promoter PgpdA, Metarhizium anisopliae, transformation

LIU Shao-fang, WANG Miao-miao, TU Xiong-bing, ZHANG Xiong-peng, NONG Xiang-qun, CAO Guang-chun, WANG Guang-jun, ZHANG Ze-hua. Two Promoters Cause Different Fluorescence Expression of egfp in Metarhizium anisopliae[J].Scientia Agricultura Sinica, 2015, 48(S): 23-31.

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Two Promoters Cause Different Fluorescence Expression of egfp in Metarhizium anisopliae

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