BCR-ABL-pIRES-SEA重组质粒在BALB/c小鼠肌肉表达分析

Expression of Reconstructed BCR-ABL-pIRES-SEA Plasmids in the Skeletal Muscles of BALB/c Mice

检测BCR-ABL-pIRES-SEA重组基因疫苗在小鼠肌肉中的表达。用已构建BCR-ABL-pIRES-SEA重组质粒免疫BALB/c小鼠一侧后肢骨骼肌,每两周一次,每次注射200μg,第七周取材用RT-PCR及免疫组织化学染色法检测目的基因在注射部位肌肉中转录和表达。结果发现:实验小鼠骨骼肌内检测到BCR-ABL和金黄色葡萄球菌肠毒素A(SEA)基因的转录,以及BCR-ABL蛋白的表达,证明所构建的BCR-ABL-pIRES-SEA重组质粒在小鼠体内可以有效地表达基因产物。

This paper is aimed to investigate the transcription and expression of BCR-ABL-pIRES-SEA fusion gene vaccines in vivo in mice. The reconstructed plasmids （BCR-ABL-pIRES-SEA） which were developed previously in our laboratory were injected into the skeletal muscles of BALB/c mice at 14d intervals for three cycles. The tran- scription and expression of BCR-ABL and staphylococcal enterotoxin A （SEA） in injection site were detected using RT-PCR and immunohistologieal methods. The BCR-ABL/SEA mRNA and protein could be identified in the iniee- tion site of BCR-ABL-plRES-SEA vaccinated mice. The reconstructed BCR-ABL-pIRES-SEA plasmids can effectively express gene production in the skeletal muscles of mice and have the common features of DNA vaccine.