[Construction of interleukin-18-PE38 fusion gene eukaryotic expression vector and its expression in the. chondrocyte].
OBJECTIVE
To construct the interleukin-18-PE38 fusion gene expression vector and explore the expression of the fusion gene in the chondrocyte.


METHODS
The recombinant eukaryotic expression vector PsecTag2B-IL-18-PE38 was constructed by inserting interleukin-18-PE38 fusion gene into eukaryotic expression vector PsecTag2B with molecular cloning technique. It was confirmed by restrictive enzymes (EcoR I) digestion assay and PCR. The vector was transfected into primary chrondrocyte by liposome protocol, and the transient expression was identified by fluorescence immunocytochemical assay.


RESULTS
Restrictive enzymes digestion analysis and PCR revealed that the interleukin-18-PE38 fusion gene was cloned into the eukaryotic expression vector PsecTag2B successfully. Immunofluorescence photograph of fluorescence immunocytochemical method confirmed that the fusion gene can be expressed in the cytomembrane and cytoplasm.


CONCLUSIONS
The results confirmed that PsecTag2B-IL-18-PE38 fusion gene can be expressed in the chondrocyte, which could serve as a foundation for the study on rheumatoid arthritis therapy.
Wu
Xia
X
Department of Biochemistry and Molecular Biology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Zhang
Lin
L
Qu
Yi
Y
Li
Ming-yuan
MY
Jiang
Zhong-hua
ZH
Li
Hong
H
D036002:ADP Ribose Transferases / Q000096:biosynthesis / Q000235:genetics*; D000818:Animals; D000831:Animals, Newborn; D001427:Bacterial Toxins / Q000096:biosynthesis / Q000235:genetics*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D005057:Eukaryotic Cells / Q000378:metabolism; D005098:Exotoxins / Q000096:biosynthesis / Q000235:genetics*; D005822:Genetic Vectors; D007155:Immunologic Factors / Q000378:metabolism; D020382:Interleukin-18 / Q000096:biosynthesis / Q000235:genetics*; D008081:Liposomes; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection; D037521:Virulence Factors / Q000096:biosynthesis / Q000235:genetics*; D000097668:Pseudomonas aeruginosa Exotoxin A
D036002:ADP Ribose Transferases; D001427:Bacterial Toxins; D005098:Exotoxins; D007155:Immunologic Factors; D020382:Interleukin-18; D008081:Liposomes; D011993:Recombinant Fusion Proteins; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A