Cloning, sequence analysis and expression of the blue-light receptor gene from the basidiomycetous mushrooms
Date Issued
2011
Date
2011
Author(s)
Gung, Chiau-Ling
Abstract
For the fruiting-body formation, there are different environmental factors that must be controlled on the edible and medical mushroom cultivation. Light is one of the most important elements influencing the fruiting-body development, which indicates that there exists photoreceptor for receiving the light stimulus in the bacidiomycetes. The bacidiomycete Corprinus cinereus, Cryptococcus neoformans, Lentinula edodes and Postia placenta have the blue-light receptors that are homologs of White Collar-1 (WC-1) in ascomycete Neurospora crassa. In this study, we attempted to clone the WC-1 homologs from the edible mushrooms: L. edodes , Flammulina velutipes, Pleurotus ostreatus, Hypsizygus marmoreus variant and Coprinus comatus. We cloned two blue-light receptor genes, photoreceptor A and photoreceptor B, from L. edodes by using specific primers and the complementary DNA template of L. edodes for PCR. The genes photoreceptor A and photoreceptor B are similar to the published blue-light receptor genes LephrA (accession no. AB279630) and LephrB (accession no. AB279630) in L. edodes. The sequence of photoreceptor A is 2,831 bp and contains intron in 56 bp. The identity of nucleotides and amino acids between photoreceptor A and LephrA are 99.5% and 99.6%. The sequence of photoreceptor B is 942 bp and the identity of nucleotides and amino acids compared to LephrB are 99.5% and 99.6%. The photoreceptor B gene is divided by four introns in the genome and the size of introns is between 52 bp and 87 bp. The photoreceptor B gene were subcloned to pET-21a(+) and expressed in Escherichia coli BL 21 (DE3). The protein product Photoreceptor B-His x6 is soluble and the molecular weight is about 37.7 kDa. In the same time, we cloned a 420 bp nucleotide from the complementary DNA of F. velutipes by using the degenerate primers. The deduced 140 amino acids of the cloned nucleotide has 28%, 28%, 24% and 31% identity to the WC-1 homologs of L. edodes, Schizophyllum commune, Phycomyces blakesleeanus and C. cinereus.
Subjects
basidiomycetous mushroom
light-sensing system
blue-light receptor
Lentinula edodes
Flammuliana velutipes
Type
thesis
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