变形链球菌表面蛋白真核表达载体pcDNA3-PAc的构建Ⅲ.变形链球菌表面蛋白PAc编码基因pac的体外扩增

Construction of streptococcus mutans surface protein antigen eukaryotic plasmid expression vector III. Amplification of the pac gene DNA fragment of srteptococcus mutans surface protein antigen by polymerase chain reaction

目的获取完整的变形链球菌表面蛋白PAc的编码基因 pac。 方法建立扩增长片段目的基因的PCR反应体系 ,根据文献报道的 pac核苷酸序列设计引物 ,使用通用缓冲液和高纯度的质粒 pPC4 1DNA为模板 ,经PCR扩增 pac基因。 结果扩增产物经琼脂糖电泳和分子量鉴定 ,为单一的 4 .7kb条带。结论成功地从质粒 pPC4 1DNA模板上扩增到全长 4 .7kb的完整 pac基因 。

Objective To obtain the complete pac gene DNA fragment of streptococcus mutans surface protein antigen.Methods Used a long-distance polymerase chain reaction (PCR) assay with plasmid pPC41 as template and primers designed by ourselves to amplify pac gene. Results A single 4.7kb DNA band was produced and estimated by gel electrophoresis. Conclusion The 4.7kb gene DNA fragment was successfully obtained from plasmid pPC41 by PCR, which could be applied in the study of nucleic acid vaccines of dental caries.