黑曲霉β-葡萄糖苷酶基因在大肠杆菌中的克隆与表达

目的构建糖类标记载体筛选克隆。方法以pPIC9K-βGl2为模板,通过PCR扩增得到全长的Bgl2基因片段,克隆至pET-28a载体,转化E.coli BL21(DE3),Cel-M9培养基筛选阳性克隆,经DNA测序分析,成功构建pET28a-Bgl2表达载体。SDS-PAGE显示,重组菌经IPTG诱导后表达了目的蛋白,其相对分子质量与预期结果相符。结果 PCR扩增得到Bgl2基因片段,SDS-PAGE显示蛋白以包涵体形式表达,克隆可用Cel-M9培养基筛选。结论用Cel-M9培养基筛选出阳性克隆为构建食品级载体奠定基础。

Cloning and expression of aspergillus niger β-Glucosidase gene in escherichia coli

Objective Building sugars marker carrier vector for screening clone.Methods Used pPIC9K-βGl2 as a template,amplified Bgl2 full-length gene fragment by PCR,cloned into pET-28a vector and transformed into E.coli BL21（DE3）,screened clones by Cel-M9 medium.The recombinant pET-28a-Bgl2 expression plasmids were successfully constructed and analyzed by DNA sequencing.From the result of the SDS-PAGE,the recombinant plasmid could express the target protein after IPTG inducment,its molecular weight met the expectation.Result Got Bgl2 gene fragments by PCR amplification,SDS-PAGE analysis showed that the protein expressed as inclusion as inclusion body,and positive clones could screened by Cel-M9 medium.Conclusion Screened clones by Cel-M9 medium and built the foundation for food-grade vectors.