地衣芽孢杆菌A13重组表达黑曲霉植酸酶phyA基因的研究

Recombinant Expression of Aspergillus niger phyA Gene in Bacillus Licheniformis A13

将黑曲霉Z6植酸酶phyA基因置于芽孢杆菌强启动子F1下,与大肠-芽孢杆菌穿梭表达载体pHY300PLK连接,重组质粒pHY300-F1-phyA电击转化地衣芽孢杆菌A13。筛选阳性克隆,获得重组菌株A13-F1-phyA,PCR扩增及酶切验证表明植酸酶基因已转入地衣芽孢杆菌。SDS-PAGE分析和表达产物的酶活性研究证明,重组菌株中植酸酶得到了有效分泌和表达。

We put phyA gene from Aspergillus niger Z6 under the strong Bacillus promoter F1 and then linked it with E.coli-Bacillus shuttle vector pHY300PLK. We also constructed the recombinant plasmid pHY300-F1-phyA and transformed it into Bacillus licheniformis A13 by electroporation. The recombinant strain A13-F1-phyA was obtained by plate screening. PCR amplification and enzyme digestion proved that phyA had been inserted into B.licheniformi.SDS-PAGE analysis and enzymatic kinetics research indicate that phytase is effectively secreted and expressed in the recombinant strain.