利用TAIL-PCR技术克隆猴头菇β-glucosidase基因

Cloning of β-glucosidase Gene by TAIL-PCR from Hericium erinaceus

根据已知的β-glucosidase基因的保守序列设计引物,从猴头菇的菌丝体中克隆到β-glucosidase基因片段B1,再利用TAIL-PCR技术扩增B1两端的侧翼序列B2和B3,B2和B3序列经Blastn、ORF和Primer5·0软件分析后,再设计B2和B3侧翼序列的特异引物对猴头菇的基因组DNA进行扩增,最终扩增到全长的β-glucosidase基因,其大小为2226bp。

A pair of special primers were designed according to the conservative sequence of the reported β-glucosidase gene. The β-glucosidase gene segment (B1) was amplified with the genomic DNA of mycelia of Hericium erinaceus and the two terminus B2 and B3 of B1 flanking sequences were amplified by TAIL-PCR. The special primers on the flanking sequence B2 and B3 were designed according to above sequences that were analyzed by using the softwares of Blastn,ORF and Primer 5.0. The full β-glucosidase gene sequence of 2228bp was then amplified with the special primers from the genomic DNA of Hericium erinaceus.