桦褐孔菌β-葡萄糖苷酶基因的克隆与定量表达分析

Cloningand Quantitative Expression of a β-Glucosidase Gene from Inonotus obliquus

利用反转录PCR(reverse transcription PCR,RT-PCR)和cDNA末端快速扩增(rapid-amplification of cDNA ends,RACE)技术从桦褐孔菌(Inonotus obliquus)菌核中克隆了β-葡萄糖苷酶基因的全长DNA序列,命名为IO-BGL。该DNA序列全长3382bp,其中开放阅读框(ORF)长度为2583bp,含有13个内含子和14个外显子,编码860个氨基酸,相对分子量为93.4kD,等电点为5.57,编码的氨基酸序列与地中海嗜蓝孢孔菌(Fomitiporia mediterranea)相似性最高(85%)。采用实时荧光定量PCR技术研究IO-BGL基因在菌核发育过程中的表达量变化,结果表明,IO-BGL基因在菌核的发育过程中,表达量呈先上升后下降趋势。

The full-length DNA sequence of a β-glucosidase gene （IO-BGL） was cloned from Inonotus obliquus sclerotia by RT-PCR and RACE. The gene had a molecular mass of 93.4 kD, a pI value of 5.57, and consisted of 3382 bp, 13 introns and 14 exons, and an open reading frame （ORF） of 2583 bp that encoded a protein of 860 amino acids. The putative amino acid sequence of IO-BGL was 85% similar to a β- glucosidase from Fomitiporia rnediterranea. Real time quantitative PCR revealed that IO-BGL expression increased during the first 170 days of sclerotium development and then decreased.