毛头鬼伞漆酶基因Cc_lac21的克隆及生物信息学和组织表达分析OA北大核心CSTPCD

To determine the'gap'sequence and possible physiological functions of the laccase gene Cc_lac21 in the genome of Coprinus comatus,the Cc_lac21 gene was cloned and analyzed by bioinformatics,and its expression level at mycelium stage and in the pileus,upper stipe and basal stipe of fruiting bodies at commercial maturation stage were detected using 18S rDNA as reference gene.The results showed that the full length of Cc_lac21 was 2 243 bp containing 12 introns and 13 exons,and the coding sequence was 1 452 bp encoding 483 amino acids.Cc_Lac21 was a stable hydrophilic protein with the first 17 amino acids at N-termi-nus as signal peptide sequence,which was secreted to extracellular space.The secondary structure of Cc_Lac21 was mainly composed of random coiled-coil and extended chain.Phosphorylation might occur on 28 serines(S),21 threonines(T)and 5 tyrosines(Y),and O-glycosylation might occur at threonine 286 and serine 381.It contained three cupredoxin protein domains including two cupredoxin super families and one CuRO_3_Tv-LCC9 of Trametes versicolor Tv-LCC like fungal laccase.It was the most closely related to laccase of Leucoagaricus leucothites and contained three motifs.Through RT-qPCR analysis,it was found that Cc_lac21 was highly expressed in the mycelium and basal stipe of matural fruiting bodies,and was lower expressed in the pileus and upper stipe in elongation growth,speculating that Cc_lac21 might participate in the lignin degradation of cultivated substrate and maturation process of stipe cell wall.The results of this study could lay a foundation for exploring the mechanism of laccase genes involved in cell wall maturation of stipe.