根据肺形侧耳(Pleurotus pulmonarius)产孢相关基因 stpp1,采用反转录 PCR(reverse transcription PCR,RT-PCR)技术克隆出糙皮侧耳(P.ostreatus)产孢相关基因同源物 stpo1全长,并对其保守域进行生物信息学分析,使用pEASY-E2原核表达系统进行该基因的原核表达,通过荧光定量PCR分析stpo1表达模式。结果表明：糙皮侧耳stpo1全长cDNA序列为2556 bp,预测编码851个氨基酸的蛋白质,具有错配修复 DNA结合域(MUTSd)、ATP 结合位点(ABC-ATPase )以及错配修复域(MutS),属于错配修复家族。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)显示stpo1在原核细胞中具有表达活性,定量 PCR 显示 stpo1在糙皮侧耳不同发育阶段显现不同的表达水平并且在成熟子实体菌褶中的表达量明显高于其它部位,表明该基因可能与糙皮侧耳的孢子形成有相关性。%Reverse transcription-PCR (RT-PCR)was used to clone a sporulation-related gene,stpo1,from Pleurotus ostreatus, and the recombinant plasmid pEASY-stpo1 was constructed and transformed into Escherichia coli BL21 (DE3).Stpo1 was 2556 bp in length,encoding 851 amino acids residues,contained MutS and ABC-ATPase domains,and belonged to the MSH4 mismatch repair family.Semi-quantitative PCR-based analysis of stpo1 expression patterns in different parts of P.ostreatus fruit bodies and at different stages of development revealed much higher levels of gene expression in the gills compared with the pileus and stipes,and a significant increase in expression at the mature fruiting body stage.
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