来源于Chaetomium cupreum的几丁质酶chi58基因克隆及特性分析

Cloning and Characteristics of Chitinase chi58 Gene from Chaetomium cupreum

以角毛壳菌cDNA文库中获得的几丁质酶基因片段(GenBank Accn:DV546055)为基础,用反向PCR技术克隆出该基因的全长cDNA序列,命名为chi58。其开放阅读框(ORF)1602bp,编码533个氨基酸组成的多肽,蛋白分子量为58kD,理论等电点为4.47。序列分析表明,该基因由2个内含子和3个外显子组成。经BlastP分析,chi58基因的氨基酸序列的保守性不高,与其相似性最高的是球毛壳菌(Chaetomium globosum XM001230073),相似性为75%。表明该基因是一条新发现的几丁质酶家族基因。该序列已收录于GenBank,登录号为EF026977,DQ886936。

The full length cDNA encoding chitinase （chi58） gene from Chaetomium cupreum was cloned through IPCR technology based on the sequence （ GenBank Accn ： DV546055） in C. cupreum cDNA library. The open reading frame was 1 602 bp encoding 533 amino acids. The protein molecular weight was 58 kD and its theoretical isoelectric point was 4.47. The sequence analysis indicates that chi58 gene contains 3 exons and 2 introns. BlastP analysis reveals that the amino acid sequences of chi58 genes from some fungal species share low similarity. The chi58 gene from C. cupreum shares the highest similarity with that from C. globosum, and the similarity is 75%. The cDNA and deduced amino acid sequence of chi58 gene were accepted by GenBank （Accession numbers： EF026977, DQ886936）.