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《Journal of Jinan University》 2006-03
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Cloning and expression of arabinosidase from Armillariella tabescens

XIE Chun-fang,HUANG Hui,YAO Dong-sheng,LIU Da-ling,LIANG Yu-qiang(Department of Bioengineering,Jinan University,Guangzhou 510632,China)  
Through random sequencing of an expression cDNA library of Armillariella tabescens,AJ620046 was detected that it has significant similarity to the arabinosidase of Bacteroides thetaiotaomicron.By using SMART-RACE technology,the full length cDNA of AJ620046 was obtained successfully.Analyzing with bioinformatics methods.Results showed the full-length cDNA has an open reading frame of 924 bp encoding a 307 amino acid protein.The full-length cDNA has significant similarity to the arabinosidase of Bacteroides thetaiotaomicron.It belongs to Arabinanase/levansucrase/invertase superfamily in 80~279 amino acid.From 58 to 184 amino acid is a typical functional domain.DNAMAN software predicted its relative molecular weight is(32 881) and pI is 4.59.The coding sequence of AJ620046 was cloned into the pPIC9 vector to create a fusion with the yeast.The pPIC9-AF recon was integrated into P.pastoris strain GS115 and expressed.The relative activity is 81.25 %.
【Fund】: 国家高技术“863”计划项目(2002AA213001);; 广东省科技攻关计划项目(2005B20601004)
【CateGory Index】: Q78
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