摘要
目的 :试图用乳酸杆菌在局部分泌干扰素 ,以达到预防和治疗病毒感染。方法 :以 pBLUE/IFNα 2b为模板 ,采用PCR技术将IFNα 2b基因扩增然后插入乳酸菌表达载体 pSC111AE ,将重组质粒电转德氏乳杆菌DM 890 9,获得重组菌株。在含乳糖的MRS培养基培养并诱导表达 ,表达产物分泌到菌液上清。结果 :分别用PCR和ELISA鉴定目的基因和表达产物 ,利用WISH VSV系统采用细胞病变抑制法 ,测定表达产物IFNα 2b的抗病毒活性 ,平均效价为 1 4 1× 10 5IU/ml。结论 :本研究为以乳酸杆菌为载体在局部产生治疗性药物奠定了基础。
Objective: To use lactobacillus as a vector to deliver interferon locally for prevention and therapy of virus infections.Methods:With pBLUE/IFN α-2b as template,the IFN α-2b gene was PCR amplified,cloned into expression vector pSC111AE,and the recombinant plasmid was electrotransformed into Lactobacillus dehrueckii Subsp.lactis DM8909.The recombinant Lactobacillus strain was grown at 37 ℃ on MRS medium containing lactose and induced for expression.Results:PCR and restrictive enzyme digestion were applied for the identification of interest gene.The supernatants were collected and tested by dot blot and ELISA for the assay of IFN α-2b.Antiviral activity was determined in cultured cells by a cytopathogenic effect(CPE) inhibition assay using WISH-VSV(vesicular stomatitis virus) system.The mean antiviral titer was 1.41×10 5 IU/ml.Conclusion:Our work provides a good basis for developing Lactobacillus as vehicles to produce therapeutics locally.
出处
《中国微生态学杂志》
CAS
CSCD
2004年第3期129-131,共3页
Chinese Journal of Microecology
基金
美国Symbigene公司资助
