摘要
目的克隆人黑色素瘤特异抗原 MAGE- 3基因 ,以制备肿瘤 DNA疫苗。方法用 RT- PCR方法制备 MAGE-3基因 ,以哺乳细胞高效表达质粒 p CI- neo为载体 ,构建重组 DNA疫苗。重组体用载体上的通用引物为测序引物 ,鉴定克隆的正确性。再将鉴定过的重组质粒用脂质体法转化 2 93细胞 ,用免疫印迹法鉴定转化细胞中 MAGE- 3基因的表达。结果正确构建了 MAGE- 3/p CI- neo重组质粒 ,并且在转化细胞中检测出了 MAGE- 3的表达。结论成功地构建了重组 MAGE- 3/p CI- neo肿瘤核酸疫苗 。
ObjectiveTo clone human melanoma antigen MAGE 3 gene to produce tumor DNA vaccine Methods Recombinant DNA vaccine was constructed by ligating MAGE 3 gene, which was prepared by RT PCR and the vector pCI neo that can express in mammals with high efficiency The recombinant was sequenced on an automatic sequencer with T7 promoter and T3 promoter sequence as sequencing primers that locate upstream and downstream of the multiple cloning site (MCS) respectively and the correctness of recombinant was determined The identified recombinant plasmids were then transformed into 293 cells with liposome, the expression of MAGE 3 was checked by western blot ResultsThe recombinant MAGE 3/pCI neo plasmid had been constructed correctly and the expression of MAGE 3 gene in transformed 293 cells had been determined ConclusionWe had constructed successfully the recombinant MAGE 3/pCI neo tumor vaccine Its anti tumor effects could be observed further in animal models
出处
《免疫学杂志》
CAS
CSCD
北大核心
2001年第6期406-409,共4页
Immunological Journal
基金
国家自然科学基金资助项目 (3990 0 134)
