采用植物叶总RNA提取试剂盒提取康宁木霉AS3.2774总RNA,根据GenBank已发表的纤维素酶CBH Ⅱ基因序列(DQ504304)设计并合成了1对特异性引物,采用RT-PCR方法扩增得到康宁木霉CBH Ⅱ基因cDNA,并成功转入大肠杆菌。序列测定结果表明,该基因序列与GenBank发表的康宁木霉CBH Ⅱ基因序列同源性达到99.86%。
The total RNA of Trichoderma koningii AS3.2774 was extracted with RNA Extraction Kit for plant leaf. A pair of primers was designed to amplify the c DNA of CBH Ⅱ gene by RT-PCR according to the published sequence of Trichoderma koningii CBH Ⅱ gene(Accession Number: DQ504304),and the obtained c DNA was cloned into E. coli and subsequently sequenced. The sequence analysis result showed that the sequence similarity between the cDNA obtained in this study and the published sequence of Trichoderma koningii CBH Ⅱ gene was 99.86%.