摘要
目的 探讨ＤＮＡ疫苗ｐＣＩ－Ｓ－ＩＲＥＳ－ＰｒｏＴα的构建及体外表达。方法 选择编码乙肝表面抗原Ｓ基因片段、内核糖体进入位点（ｉｎｔｅｒｎａｌ ｒｉｂｏｓｏｍｅ ｅｎｔｒｙ ｓｉｔｅ，ＩＲＥＳ）片段和胸腺素ａ原（ｐｒｏｔｈｙｍｏｓｉｎ α，ＰｒｏＴα）基因片段，利用基因重组技术构建成ＤＮＡ质粒ｐＣＩ－Ｓ－ＩＲＥＳ－ＰｒｏＴα，并将该ＤＮＡ质粒转染成纤维细胞ＮＩＨ３Ｔ３。结果 不仅可通过ＲＴ－ＰＣＲ检测到其ＲＮＡ的表达，而且ＥＬＩＳＡ也检测到其抗原表达。结论 构建的质粒可作为ＤＮＡ疫苗，进一步用于免疫动物。
Objective To investigate the construction of DNA vaccine pCI - S - IRES - ProTα and its in vitro expression. Methods The DNA plasmid pCI - S - IRES - ProTα, which contained the S gene encoding surface antigen of HBV, internal ribosome entry site and cytokine prothymosin α gene, was constructed by gene technology. The plasmid PCI - S - IRES - ProTa was transferred into the cell line NIH3T3. The in vitro expression of DNA plasmid was observed. Results Transcripts of the plas- mid were detected by RT - PCR,and HBsAg was also detected by ELISA. Conclusion The plasmid pCI - S - IRES - ProTa can be used as a DNA vaccine.
出处
《上海第二医科大学学报》
CSCD
2001年第3期193-195,共3页
Acta Universitatis Medicinalis Secondae Shanghai
基金
国家自然科学基金资助项目(39770680)
上海市博士点基金资助项目(98
99年度)
