水稻膨胀素基因OsEXP2在毕赤酵母中的表达

膨胀素是一类可以诱导失活细胞壁伸展的蛋白质,试验通过克隆水稻膨胀素(OsEXP2)基因并在巴斯德毕赤酵母中表达,以研究其对纤维素的作用。根据已知序列设计引物,经RT-PCR克隆OsEXP2基因,并插入毕赤酵母表达载体pGAPHα中,位于α-信号肽下游,得到重组质粒pGAPHα-OsEXP2。将其线性化后通过电转化,转化毕赤酵母菌株GS115,OsEXP2基因通过同源重组整合到毕赤酵母的基因组上,得到重组菌株GS115/pGAPHα-O-sEXP2。该菌株发酵液可使滤纸发生崩解现象,并使纤维素酶滤纸酶活性(Filter Paper Enzyme Activity,FPA)提高了47.87%。

Expression of α-Expansin2 gene OsEXP2 from rice in Pichia pastoris

Expansins are proteins that mediate extension of inactivated cell walls. The α-Expansin2 gene from rice (Oryza sativa. L) was cloned and expressed in Pichia pastoris, to study the effect of it on cellulose. The OsEXP2 gene was cloned by RT-PCR with primers designed according to known the sequence, and the product was ligated with expression vector pGAPHα, downstream of α-factor signal peptide sequence, getting a recombinant plasmid pGAPHα-OsEXP2. It was linearized and transformed into P.pastoris GS115 with the method of electrotransformation, and the OsEXP2 gene was integrated into the P.pastoris genome through homologous recombination. Generated recombinant strain GS115/ pGAPH -αOsEXP2. It was proved that the fermentation broth of the strain made filter paper disintegrated, and improved cellulase FPA by 47.87%.