Cloning And Characteriztion Of The Blue Light Photoreceptor Pcry From Pleurotus Ostreatus

Light is the primary environmental signal, through which does the biology receive the light signal, affecting the growth process of the organism. The research on light receptor in Pleurotus ostreatus which is an important edible fungus is very limited, because the light receptor gene associated with Pleurotus ostreatus has not been cloned. Therefore, the 3 ’and 5’ terminal fragment about the Pcry was cloned through RACE and Nest PCR, using total RNA which was extracted from Pleurotus ostreatus New 831 as template in this study. The ful- length c DNA was secured by PCR which was used the sequencing of 3 ’and 5’ ends as the primer-template, meanwhile the correctness and the function of the gene was preliminary verified by semi-quantitative PCR and biological information methodology. The total goal of this experiment was to obtain the mechanism of Pleurotus ostreatus photomorphogenesis, while it also can lay the foundation with the blue how to influence the growth and development of Pleurotus ostreatus and further study about the gene function.The main results are obtained as follows:(1) The analysis showed that the ful- length c DNA was 1725 bp, of which the open reading frame(ORF) was 1572 bp, encoding 523 amino acids. Bioinformatics analysis predicted that this gene belonged to the family of cryptochrome and the protein was an optical signal protein, so the gene was named Pcry(Pleurotus ostreatus blue light receptor cryptochrome). Bioinformatics analysis also demonstated that the protein can regulate transcription and participate in signal transduction. The molecular weight of the protein was 57.44 KDa, and the isoelectric point was 4.9.(2) The results of semi-quantitative PCR indicated that the gene(Pcry) expression was the highest in fruiting bodies, but the expression was the lowest in mycelia at different growth period. The gene(Pcry) expression was highest after blue light irradiation in mycelium, while the gene(Pcry) expression was weakest in the dark. The results demonstated that the gene had an influence on the growth process of Pleurotus ostreatus. The expression was increased significantly after blue light irradiation, indicating that this gene may increase the transcription in the blue light stimulation. It proofed that the gene might receive a blue signal and take part in gene transcription which was mediated by theblue light-receptor.(3) The open reading frame(ORF) of the gene was verified by p EASY-E2 expression system, and the results declared that the gene could be express in prokaryotic cel s.