FimA plasmid construction and protein purification of Porphyromonas gingivalis
10.3969/j.issn.1001-3733.2018.02.005
- VernacularTitle:牙龈卟啉单胞菌菌毛质粒构建和蛋白纯化
- Author:
Jiangman CHEN
1
;
Yonggang LI
;
Xin LI
Author Information
1. 锦州医科大学附属第二医院
- Keywords:
Porphyromonas gingivalis;
Fimbriae(FimA);
Fusion protein
- From:
Journal of Practical Stomatology
2018;34(2):168-171
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To construct gine expression plasmid and to purify the protein of fimbriae(FimA) of Porphyromonas gingivalis (P. gingivalis). Methods: The bacteria DNA was extracted from P. gingivalis by using commercial kit. FimA gene was cloned after PCR. Plasmid was constructed and transformed into competent cells. Optimal conditions of protein induction were chosen. The fusionprotein was purified by Glutathione S-Transferase affinity chromatography. Fusion protein was confirmed by Western blot method. Results: The highest protein expression by the constructed plasmid was obtained at the low temperature and high concentration after 12 hours induction. The protein was confirmed by Western blot. Conclusion: A highly purified P. gingivalis FimA protein was successfully obtained.
