Cloning and Functional Analysis of hat Promoter from Cordyceps militaris
In this study,a promoter region of hat gene(Histone acetyltransferase)with a length of 1 509 bp was cloned from Cordyceps militaris.Sequences and structure were analyzed by using the PlantCARE database.Results showed that the hat promoter carried those typical cis-acting regulatory elements,such as CAAT-box and TATA-box.It also contained the G-box which was a cis-acting regulatory element involving in the responsibility to light,the SARE which was also a cis-acting element known to involve in the responsibility to salicylic acid and so on.A GFP(Green fluorescent protein)reporter cassette carrying the hat promoter,gfp gene and gpd terminator was successfully constructed for analysis.After introduction of the cassette into C.militaris through the ATMT method,transformants were applied for quality check with PCR and qRT-PCR techniques,and examinations of green fluorescence as well.As a result,a strong green fluorescence was detected in the mycelium of C.militaris.The present study thus provides us with a fundamental basis for the reconstruction of endogenous promoter elements available for transformation experiments in edible fungi and establishment of a highly effective expression system of heterologous genes in C.militaris.
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