猪圆环三型病毒抗原蛋白表达体系的优化探究

Optimization of Porcine Circovirus Antigen Protein Expression System and Screening of Superior Monoclonal Strains

目的:探究PCV3型病毒抗原蛋白表达的最优条件。方法:在摇瓶表达的过程中,通过控制变量异丙基硫代半乳糖苷(IPTG)的浓度、诱导时菌液的浑浊度(OD600)的大小条件对比PCV3型病毒抗原蛋白表达的效果。结果:在转速、补料速度、诱导后温度相同的情况下,分别设三个条件过夜诱导表达后,OD600≈1.0,加入IPTG浓度为100μL的表达量最高,三个蛋白的表达量也最均一,这是目前所有尝试后最好的且最适合PCV3表达的条件。结论:本实验中,PCV3型病毒抗原蛋白表达的最优条件为IPTG浓度为100μL、菌液浑浊度1.0。

Objective:Toexplore the optimal conditions for the expression of antigen protein of PCV3 virus.Methods : The concentration of isopropylthiogalactoside (IPTG) was changed by controlling variable method during shaking flask, and the turbidity (OD600) of bacterial solution was induced. Results: under the same conditions of rotating speed, feeding speed and temperature after induction, the expression of OD600 ≈ 1.0, under three conditions of overnight induction, the expression of PCV3 virus antigen protein was compared with that of PCV3 virus antigen protein expression. The expression level of IPTG 100 μL was the highest and the expression of three proteins was the most uniform, so this was the best condition for PCV3 expression after all attempts. Conclusion: When the concentration of IPTG was changed, the size of OD600 was induced. The optimal conditions for the expression of PCV3 virus antigen protein were found out among the three conditions of the duration of shaking flask after induction.