[Expression of receptor-associated protein of the Heymann nephritis pathogenic epitope in Escherichia coli].
To study the pathological phenotype of pathogen and the mechanism of the immunocomplex in Heymann nephritis, we utilised DNA recombinant technique, to construct plasmid pGEX-R93 by inserting the cDNA encoding of receptor-associated protein of Heymann nephritis (HN) pathogenic epitope into the pGEX. The pGEX-R93 coding region fragment was confirmed by enzyme cutting and sequencing analysis. After the pGEX-R93 was transfected into DH5 alpha, the expression of GST-RAP fusion protein was induced by isopropylthio-beta-D-galactoside are the overproduced protein consituted 39.4% of the total cellular protein. Western blot analysis showed that the antiserum for GST-RAP recognized the 44,000 alpha protein band of natural antigen in HN. In addition, immunofluorescence analysis showed that the antiserum also distinctively recognized the ciliated membrane antigen of the proximal convoluted tubule of the rat kidney.
Hu
Y
Y
Department of Molecular Biology, Nantong Medical College, Jiangsu.
Zhang
W
W
Zhu
A
A
D000818:Animals; D000939:Epitopes / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005921:Glomerulonephritis / Q000276:immunology; D026541:Heymann Nephritis Antigenic Complex; D007106:Immune Sera; D007687:Kidney Tubules, Proximal / Q000276:immunology; D008562:Membrane Glycoproteins / Q000378:metabolism*; D010957:Plasmids; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology; D014162:Transfection
D000939:Epitopes; D026541:Heymann Nephritis Antigenic Complex; D007106:Immune Sera; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins