ISSN:
1617-4623
Keywords:
Key wordsLentinus edodes
;
Glyceraldehyde-3-phosphate dehydrogenase gene
;
Restriction enzyme-mediated integration (REMI)
;
Hygromycin resistance
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract To construct a vector for high-level expression of heterologous genes in Lentinus edodes, the L. edodes GPD promoter, which is expressed constitutively and strongly, was fused to a hygromycin B phosphotransferase gene (hph) derived from Escherichia coli as a selective marker. Using the resulting pLG-hph construct, L. edodes was efficiently transformed to hygromycin resistance by restriction enzyme-mediated integration (REMI). The restriction enzyme concentrations yielding the maximal numbers of transformants by the REMI method were 10 U per transformation in the case of BglII and 25–50 U in the case of HindIII. Southern analysis of the transformants indicated that some 50% of plasmid integrations were REMI-mediated events. These results indicate that pLG is a useful vector for transformation of L. edodes. Deletion analysis of the GPD promoter region suggested that the segment between positions −442 bp and −270 bp relative to the transcription start point may be essential for function.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/s004380050033
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