摘要
根据已知的内切葡聚糖酶和β-葡萄糖苷酶基因ORF全长序列,分别设计1对带有粘性末端酶切位点EcoRI和NotI的特异性引物,PCR克隆得到桦褐孔菌内切葡聚糖酶(endo-l,4-β-D-glucanase,EG)和β-葡萄糖苷酶(beta-glucosidase,BGL)ORF基因全长,将酶切后的目的片段分别与真核表达载体pPICZαA连接,构建出真核表达载体pPICZαA-EG和pPICZαA-BGL,并成功转化到毕赤酵母GS115中,为下一步纤维素酶的表达和功能验证奠定了基础。
A pair of specific primers with viscous terminal cleavage sites EcoRI and NotI were designed according to the known full length sequence of endo-l,4-β-D-glucanase and beta-glucosidase gene ORF.PCR was used to clone the full length of ORF gene of endo-l,4-β-D-glucanase and beta-glucosidase.The target fragments were ligated with the eukaryotic expression vector pPICZαA respectively.The eukaryotic expression vectors pPICZαA-EG and pPICZαA-BGL were constructed and ligated into the Pichia pastoris GS115,which laid the foundation for the next step in cellulase expression and functional validation.
出处
《延边大学农学学报》
2017年第3期1-6,18,共7页
Agricultural Science Journal of Yanbian University
基金
国家自然科学基金项目(31160408和31460536)
关键词
桦褐孔菌
内切葡聚糖酶
Β-葡萄糖苷酶
真核
载体构建
Inonotus obliquus
endo-l,4-β-D-glucanase
beta-glucosidase
eukaryotic
vector construction
