[Cloning of the RgpAcd gene of Porphyromonas gingivalis and its expression in E. coli].
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.


CONCLUSIONS
The protein of RgpAcd will be obtained for further study and its protein was correctly expressed in E. coli BL21 cells.
Xu
Jing
J
Dept. of Stomatology, The Sixth People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai 200233, China.
Li
Ang
A
Gou
Jian-Zhong
JZ
Xu
Yuan-Chao
YC
Rao
Guo-Zhou
GZ
Liu
Zheng
Z
Xie
Hong-Guo
HG
D003001:Cloning, Molecular; D019976:Cloning, Organism; D004926:Escherichia coli; D005822:Genetic Vectors; D016133:Polymerase Chain Reaction; D016966:Porphyromonas gingivalis; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins
D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins