地衣芽孢杆菌A13重组表达黑曲霉植酸酶phyA基因的研究

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。

Recombinant Expression of Aspergillus niger phyA Gene in Bacillus Licheniformis A13

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.