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  • Title: [Cloning of the RgpAcd gene of Porphyromonas gingivalis and its expression in E. coli].
    Author: Xu J, Li A, Gou JZ, Xu YC, Rao GZ, Liu Z, Xie HG.
    Journal: Hua Xi Kou Qiang Yi Xue Za Zhi; 2006 Oct; 24(5):400-3. PubMed ID: 17315645.
    Abstract:
    OBJECTIVE: To clone the catalytic domain gene sequence of RgpAcd of Porphyromonas gingivalis (P. gingivalis) and to induce its fusion expression in E. coli. METHODS: The desired DNA fragment RgpAcd was obtained by PCR and was separately sequenced and identified by inserting into inter-vector pMD18-T vector. The correctly fragment was linked with and cloned into a prokaryotic expression vector pET-15b. The recombinant expression plasmid which had been confirmed by enzymes digestion was transformed to E. coli competent cells BL21 (DE3) and expression of fusion protein was induced by IPTG. RESULTS: A 1 476 bp specific fragment was obtained and DNA sequencing showed that the fragment was consistent with those of the published. After induction with IPTG, a fusion protein of 5 x 10(4) was visualized on SDS-PAGE gel. CONCLUSION: The protein of RgpAcd will be obtained for further study and its protein was correctly expressed in E. coli BL21 cells.
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