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目的 :构建中国人前列腺特异膜抗原 (PSMA)基因编码区序列的真核表达载体并进行序列测定。方法 :从前列腺癌组织中提取总RNA ,采用RT -PCR技术扩增前列腺特异膜抗原的基因编码区序列 ,将序列定向克隆至真核表达载体pcDNA3 0 ,并进行序列测定。结果 :本实验所扩增的中国人PSMA基因编码区序列与Israeli等报道的序列同源性为 99 7% ,目的基因与载体正确连接。结论 :成功克隆了PSMA编码区序列 ,并构建了其真核表达载体 ,为PSMA基因修饰的树突状细胞疫苗的前列腺癌生物治疗奠定基础。
OBJECTIVE: To construct a eukaryotic expression vector encoding the coding region of Chinese prostate specific antigen (PSMA) gene and to determine its sequence. Methods: Total RNA was extracted from prostate cancer tissues. The gene coding region of prostate specific membrane antigen was amplified by RT-PCR. The sequence was cloned into eukaryotic expression vector pcDNA3.0 and sequenced. RESULTS: The sequence of the coding region of Chinese PSMA amplified by this experiment was 99.7% homologous to that reported by Israeli et al. The target gene was correctly linked with the vector. CONCLUSION: The sequence of PSMA coding region was successfully cloned and its eukaryotic expression vector was constructed, which laid the foundation for the biological treatment of prostate cancer with PSMA gene modified dendritic cell vaccine.