牙龈卟啉菌牙龈蛋白酶K催化结构域的克隆及其在大肠杆菌中的表达

Cloning of the Kgpcd of Porphyromonas Gingivalis and its Expression in E.coli.

目的 :克隆牙龈卟啉菌蛋白酶K催化结构域 (kgpcd)基因 ,并使其在大肠杆菌中获得表达。方法 :利用PCR方法克隆kgpcd ,并用基因重组融合表达技术获得在大肠杆菌中的表达。结果 :克隆基因测序结果与GeneBank数据库中的序列一致 ,经诱导表达见Mr为 5 6× 1 0 3 的融合蛋白。结论 :成功克隆了kgpcd的基因 ,并在大肠杆菌中表达了KGPcd蛋白 。

Objective:To clone the catalytic domain of gingipain K (kgpcd) of Porphyromonas gingivalis (Pg) and expression of KGPcd fusion protein in Escherichia coli.Methods:The desired DNA fragment kgpcd was obtained by PCR and its expression was used by contruction of fusion protein technique.Results:A 1467bp specific fragment was obtained and DNA sequencing showed that the fragment was consistent with those of the published. After induction with IPTG, a new 56kDa protein appeared on SDS-PAGE gel.Conclusions:The kgpcd of Pg was cloned successfully and its protein was expressed in E.coli BL21.