Construction of eukaryotic expression vector for KGPcd gene from Porphyromonas gingivalis and expression in mammalian cells

This study aimed at constructing secretory eukaryotic expression vector of KGPcd gene encoding whole amino acid residues of mature KGPcd from Porphyromonas gingivalis and investigating the transcription and expression of recombined plasmid VR1020/KGPcd in mammalian cells. Eukaryotic expression plasm...

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Bibliographic Details
Published inHua xi kou qiang yi xue za zhi Vol. 22; no. 6; p. 503
Main Authors Zhang, Feng-qiu, Yang, Lian-jia, Wu, Zhi-fen, Qin, Hong-yan
Format Journal Article
LanguageChinese
Published China 01.12.2004
Subjects
Animals
Bacterial Proteins - genetics
Cercopithecus aethiops
COS Cells
Cysteine Endopeptidases - genetics
Genetic Vectors
Plasmids
Porphyromonas gingivalis - genetics
Transfection
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Summary:This study aimed at constructing secretory eukaryotic expression vector of KGPcd gene encoding whole amino acid residues of mature KGPcd from Porphyromonas gingivalis and investigating the transcription and expression of recombined plasmid VR1020/KGPcd in mammalian cells. Eukaryotic expression plasmid VR1020/KCPcd was constructed by using molecular cloning methods. Then, the VR1020/KGPcd was transfected into mammalian cell COS7 with Lipofectamine 2000 according to the manufacturer's instruction. The transcription of VR1020/KGPcd was assayed by reverse transcription polymerase chain reaction (RT-PCR). The expression product of VR1020/KGPcd was analyzed by using indirect immunofluorescence. The protein secretion in cultural medium was detected by ELISA method. It proved that the VR1020/KGPcd could be transcribed and translated into transfected COS7 cells. The expressed targeted protein could be secreted into cultural supernatant and could be detected by ELISA. The eukaryotic expression plasmid of VR1020/KGPcd w
ISSN:1000-1182