中性β-葡萄糖苷酶基因表达载体的构建

Constrcuting Expression Vector of Neutral Cellulase β-glucosidase

利用RT-PCR方法从芽孢杆菌Bacillus Subtilis CY110中克隆出β-葡萄糖苷酶基因,将此目的基因连接到pBS-T载体后导入大肠杆菌DH5α。测序结果表明,获得的目的基因片段大小为1 398 bp,该序列与Gen-Bank中的β-葡萄糖苷酶基因ABQN01000006.1序列相比同源性达到99.8%,氨基酸序列同源性达到99.5%。将产物与pET-28 a表达载体连接,经双酶切检验,证明原核表达载体构建成功。

The β-glucosidase gene was cloned from Bacillus Subtilis CY110 genome with RT-PCR methods. Connect the target gene to pBS-T carrier and put the transformant into E. coll. The results of sequence showed that the cloned gene fragment was 1398bp. Compared the sequence with the β-glucosidase gene in Genbank, the homology of nucleic acid sequence and amino acid sequence could reach 99.8% ,99.5%. Connect pBS-T/ β-glucosidase to Plasmid pET-28a ,it was proved the pET-28a/β-glucosidase was constructed successfully by double digests.