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

Jing Xu; Ang Li; Jian Zhong Gou; Yuan Chao Xu; Guo Zhou Rao; Zheng Liu; Hong Guo Xie

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

Jing Xu
, Ang Li
, Jian Zhong Gou
, Yuan Chao Xu
, Guo Zhou Rao
, Zheng Liu
, Hong Guo Xie

医学部

Shanghai Jiao Tong University

科研成果: 期刊稿件 › 文章 › 同行评审

摘要

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.

源语言	英语
页（从-至）	400-403
页数	4
期刊	Hua Xi Kou Qiang Yi Xue Za Zhi / West China Journal of Stomatology
卷	24
期	5
出版状态	已出版 - 1 10月 2006

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@article{32f59453131b4abebe22bb32cc67f95d,

title = "Cloning of the RgpAcd gene of Porphyromonas gingivalis and its expression in E. coli",

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.",

author = "Jing Xu and Ang Li and Gou, \{Jian Zhong\} and Xu, \{Yuan Chao\} and Rao, \{Guo Zhou\} and Zheng Liu and Xie, \{Hong Guo\}",

year = "2006",

month = oct,

day = "1",

language = "英语",

volume = "24",

pages = "400--403",

journal = "Hua Xi Kou Qiang Yi Xue Za Zhi / West China Journal of Stomatology",

issn = "1000-1182",

publisher = "Editorial Department of West China Journal of Stomatology",

number = "5",

}

TY - JOUR

T1 - Cloning of the RgpAcd gene of Porphyromonas gingivalis and its expression in E. coli

AU - Xu, Jing

AU - Li, Ang

AU - Gou, Jian Zhong

AU - Xu, Yuan Chao

AU - Rao, Guo Zhou

AU - Liu, Zheng

AU - Xie, Hong Guo

PY - 2006/10/1

Y1 - 2006/10/1

N2 - 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.

AB - 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.

UR - https://www.scopus.com/pages/publications/84930399836

M3 - 文章

C2 - 17315645

AN - SCOPUS:84930399836

SN - 1000-1182

VL - 24

SP - 400

EP - 403

JO - Hua Xi Kou Qiang Yi Xue Za Zhi / West China Journal of Stomatology

JF - Hua Xi Kou Qiang Yi Xue Za Zhi / West China Journal of Stomatology

IS - 5

ER -

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

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