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흰쥐에서 Glomerular Basement Membrane Heparan Sulfate Proteoglycan(GBM HSPG)의 클론
고철우,Kasinath, B . S . 대한신장학회 1996 Kidney Research and Clinical Practice Vol.15 No.3
Although full length cDNA sequences of murine and human basement membrane HSPG are known, rat basement membrane HSPG has not been cloned. Oligonucleotide primers were synthesized from the published sequences of domainI of mouse and human basement membrane HSPG, perlecan, where the homology is 9996. The primers were employed in RT-PCR using polyARNA from rat glomerular epithelial cells (GEC) in culture to obtain a 497-bp product, rat perlecan domain-I (RPD-I). RPD-I was sequenced by dideoxy chain termination method using either the M-13 primers or the perlecan domain-I specific primers. The deduced amino acid sequence of the rat product showed 85 % and 88 % homology to the published sequences of domain-I of mouse and human perlecan, respectively. In RPD-I, the three sites for heparan sulfate glycosaminoglycan attachment with the consensus sequence SGD and the sequence NFT for N-linked oligosaccharide attachment are fully conserved, similar to mouse and human species. By northem analysis, employing total and polyA'RNA of rat GEC, RPD-I detected a single large (-12 kb) transcript, similar in size to rat glomerular perlecan reported previously. Our data demonstrate that GBM HSPG synthesized by rat GEC shares similarity with murine and human perlecan basement membrane HSPG.