金龟子绿僵菌酸性海藻糖酶基因的克隆及其序列特征分析

根据酸性海藻糖酶的蛋白质序列设计引物,PCR扩增出CQMa102 ATM1的cDNA和DNA序列,登录号分别为:DQ237957,EF190950.序列分析表明,ATM1 DNA序列含有3个内含子,其开放阅读框(ORF)编码1个含1 073个氨基酸的蛋白质,具有1个20个氨基酸的信号肽序列和含有30个可能的N-糖基化位点(Asn-Xaa-Ser/Thr).NCBI序列比对(Blastn)显示该蛋白与Aspergillus fumigatus的alpha、alpha-trehalose glucohydrolase,Aspergillus nidulans的酸性海藻糖酶前体和Talaromyces emrsonii的酸性海藻糖酶分别有62%、59%和57%的氨基酸相似性,与另外两个真菌Saccharomyces cerevisiae(Ath1p)和Candida albi-cans(Atc1p)的酸性海藻糖酶也具有约25%的氨基酸相似性.Southern杂交表明,ATM1基因在CQMa102基因组中为单拷贝.

Cloning and Characterization of the Acid Trehalase Gene in Metarhizium anisopliae CQMa102

Based on the known amino acid sequence of the acid trehalase in Metarhizium anisopliae CQMa102,the whole cDNA sequence and its whole genomic DNA of ATM1 gene in M.anisopliae CQMa102 was obtained(GenBank accession:DQ237957,EF190950).The sequence analysis shows that the DNA sequence includes three introns,and the cDNA encodes a protein of 1 073 amino acid residues,which has a 20 aa signal peptide sequence and 30 putative N-glycosylation sites.Comparison of the amino acid sequence with the sequences found in protein data bases using the NCBI Blastn algorithm revealed the protein shared 62%,59% and 57% similarities with alpha,alpha-trehalose glucohydrolase in Aspergillus funigatus, acid trehalase precursor in Aspergillus nidalans and acid trehalase in Talaromyces emrsonii,respectively.This protein also shared about 25% similarities with the acid trehalases from Saccharomyces cerevisiae(Ath1p) and Candida albicans (Atc1p).Southern analysis indicated that ATM1 gene was present as a single copy in this M.anisopliae strain.