口服基因疫苗质粒稳定性测定

在研究口服基因疫苗的过程中 ,为确保口服疫苗剂量 ,对转化细菌中质粒稳定性的动态变化 ,细菌浓度OD60 0 值与有效细菌 (携带质粒的细菌 )的关系等进行了测定 ,结果显示PAR3132在培养 10h前 ,pcDNA3 VP1在培养 6h前质粒在细菌中的稳定性可维持在 95 %以上 ,但当培养时间超过 16h ,2种质粒的稳定性在Amp为 5 0g/L时均不足 2 0 %。高浓度Amp可提高质粒稳定性。在质粒稳定性维持于 95 %的时段内 ,细菌浓度的OD60 0 值与有效细菌的量呈线性关系。因此 ,在确定口服基因疫苗剂量时 ,应根据细菌各自的情况决定振摇的时间和采用的OD值。

Study on the Kinetic Changes of Stability of the Plasmids in the Transformed Bacteria as Orally Administered Gene Vaccine

In order to determine the dose of vaccine in the course of study of orally administered gene vaccine, we detected the kinetic changes of stability of the plasmids in the transformed bacteria and the relation between the value OD-{600} of bacterial concentration and the bacteria that carry the plasmid. The results showed that the stability of plasmid in the bacteria maintains above 95 % in 10 hours of culture for PAR3132 and in 6 hours of culture for pcDNA3-VP1.When the culture time passes 16 hours, both the stabilities in the medium supplemented with 50 mg/mL Ampcillin decreased to less than 20 %. Higher concentration of Ampicillin may help to increase the plasmid stability. During the period that the plasmid stability is above 95 %, there is a linear relation between the value OD-{600} of bacterial concentration and the bacteria that carry the plasmid. It is suggested that the determination of dose of orally administered gene vaccine dependents on the culture time and the selected OD600 value.;