聚γ谷氨酸生物合成与DegR关系及其免疫佐剂效果研究

发布时间：2018-03-19 10:31

  本文选题：地衣芽胞杆菌　切入点：聚γ谷氨酸　出处：《动物医学进展》2017年04期 　论文类型：期刊论文

【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。
[Abstract]:In order to investigate the effect of DegR on the biosynthesis yield of poly 纬 -glutamic acid (纬 -PGA), the adjuvant effects of 纬 -glutamic acid adjuvant and traditional aluminum adjuvant were compared. The shuttle expression vector pHY-degRwas constructed by using P43 promoter, and the recombinant strain WX-02 / pHYdegRwas obtained by electrotransformation of Bacillus licheniformis WX-02. The protein vaccine of Staphylococcus aureus inactivated by formalin was also used as immunogen. The related vaccines were prepared with different molecular weight poly (纬 -glutamic acid) adjuvant and aluminum adjuvant. Staphylococcus aureus was injected into mice for 7 days after immunization. The relative immune protection rate was calculated according to the number of mice killed. The results showed that, after overexpression of degR, the relative immune protection rate was calculated. The yield of polyglutamic acid biosynthesis was increased to 28.5g / L, which was 22.6g / L higher than that of the control strain WX-02/pHY300. The molecular weight of 2 脳 10 ~ 3 ku poly (纬 -glutamic acid) as adjuvant had the best relative immune protection rate of 78.9%, which was better than that of aluminum adjuvant (52.6%) and no adjuvant was added. The immune protection rate was 42.1%. The results showed that. Overexpression of degR gene can improve the biosynthesis of poly (纬 -glutamate), and polyglutamic acid as a vaccine adjuvant can enhance the immune effect of the vaccine and can be used as an ideal vaccine adjuvant.
【作者单位】： 长江大学动物科学学院;长江大学城市建设学院;
【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)
【分类号】：S852.4
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本文编号：1633883