聚γ谷氨酸生物合成与DegR关系及其免疫佐剂效果研究  

Study on Poly-γ-glutamic Acid Biosynthesis with degR Overexpression and Poly-γ-glutamic Acid Adjuvant Effect

作　　者：田光明[1] 游蕾[2] 

机构地区：[1]长江大学动物科学学院,湖北荆州434023 [2]长江大学城市建设学院,湖北荆州434023

出　　处：《动物医学进展》2017年第4期46-51,共6页Progress In Veterinary Medicine

基　　金：湖北省教育厅青年人才基金项目(q20151306);湿地生态与农业利用教育部工程研究中心基金项目(KF201509)

摘　　要：为探讨调节因子DegR对聚γ谷氨酸(γ-PGA)生物合成产量的影响,比较聚γ谷氨酸佐剂和传统铝佐剂的辅助免疫效果,利用P43启动子构建了穿梭表达载体pHY-degR,通过电转化地衣芽胞杆菌WX-02获得重组菌株WX-02/pHYdegR。同时以福尔马林灭活的金黄色葡萄球菌蛋白疫苗作为免疫原,分别以不同分子质量聚γ谷氨酸佐剂和铝佐剂制备相关疫苗并免疫小鼠,7d后注射金黄色葡萄球菌,根据小鼠死亡数,计算其相对免疫保护率。结果显示,过表达degR后,聚γ谷氨酸生物合成产量提高至28.5g/L,相比对照菌株WX-02/pHY300提高22.6%。分子质量2×10~3 ku聚γ谷氨酸作为佐剂的相对免疫保护率效果最好(78.9%),优于铝佐剂免疫保护率(52.6%)和未添加佐剂免疫保护率(42.1%)。结果表明,过表达degR基因能提高聚γ谷氨酸生物合成量,且聚γ谷氨酸作为疫苗佐剂能增强疫苗免疫效果,可作为理想的疫苗佐剂。To investigate the effect of the overexpression of degR gene on the yield oi poly-γ-glutamlc aclct and the immune effect of γ-PGA adiuvant, an engineered strain of B. licheniformis （WX-02/pHYdegR） was developed. Transformation of B. licheniformis WX-02 with a shuttle plasmid pHY-degR builded by strong promoter P43 was performed via electroporation. Meanwhile,formalin-killed Staphylococcus aureus proteins were used as immunogen with different adjuvants （alum adiuvant and γ-PGA adjuvant） to immunize healthy mice. The relative immune protection rate was measured after γ days according to the mouse mortality after injecting viable Staphylococcus aureus. The results showed that the yield of γ-PGA reached 28.5 g/L,increased by 22.6% in comparison with that of the control （23.25 g/L） by enhancing expression of degR gene in vivo. In the immune-protective experiment, the immunogen mixed with γ-PGA adjuvant （2X10^3 ku） had the best immune effect （78.9%） compared with alum adjuvant （52.6%） and pure immunogen （42.1%）. Therefore,γ-PGA adjuvant can be used as an ideal adjuvant in veterinary field.

关 键 词：地衣芽胞杆菌 聚γ谷氨酸 degR 疫苗佐剂 免疫效果 

分 类 号：S852.4[农业科学—基础兽医学]