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目的旨在构建一种新型的表面蛋白防龋疫苗—DNA防龋疫苗。方法从表面蛋白pac基因上利用BamHI和Sacl双酶切，获得一个3281bp的片段。将该片段定向插入真核表达载体PSVL，再将连接后新构成的质粒pSVL／pac转入氯化钙法制备的感受态大肠杆菌E．coliDH5a。用BamHI和SacI双酶切，琼脂糖电泳证实pSVL／pac构建的正确性。结果用定向克隆的方法获得了一个含pac基因的真核表达质粒pSVL／pac即表面蛋白DNA疫苗。结论利用定向克隆和氯化钙法可以较为简便和快速地构建表面蛋白的DNA防龋疫苗。
The purpose is to construct a new type of surface protein anti-caries vaccine-DNA anti-caries vaccine. Methods A 3281bp fragment was obtained by digestion with BamHI and Sacl from the surface protein pac gene. The fragment was inserted into the eukaryotic expression vector PSVL, and then the newly constructed plasmid pSVL / pac was ligated into the competent E. coli E prepared by calcium chloride method. coliDH5a. Restriction enzyme digestion with BamHI and SacI confirmed the correctness of pSVL / pac construction by agarose gel electrophoresis. Results An eukaryotic expression plasmid pSVL / pac, a surface protein DNA vaccine containing the pac gene, was obtained by directional cloning. Conclusion The use of directional cloning and calcium chloride method can be more simple and rapid construction of surface protein anti-caries DNA vaccine.