Abstract

We have cloned the 250kb area centromeric of the HLA class II region which corresponds to the mouse T/t complex region by YAC and cosmid vectors. were identified, ----RING1----COLLIIA2-DPB2 from the centromeric side. A cosmid clone (40 kb insert) containing HKE6 and other several CpG islands with rare cutting enzyme recognition sites (Not I, BssH II, Eag I, Sac II) was subjected to nucleotide sequence determination. Sheared DNA has been cloned into the pUC19 vector and the nucleotide sequences were determined using automated DNA sequencer (ABI 373 and 377). A total of 780 clone sequences (including forward and reverse sequences) were edited mannually and assembled using two softwares (Sequencher ver.3.0 and ATSQ programs). Primers were designed from the ends of the contigs to perform PCR, in some case followed by Southern hybridization, to order the contigs and cover the gaps. Five exons of the RING1 gene, 7 exons of HKE6 and other potential coding exons were identified and characterized. The HKE6 gene is a human homologue of the mouse Ke6 gene which might be involved in the manifestation of polycystic kidney disease. Identification and characterization of non-MHC genes around the MHC region may provide insights into physical and gene organization of human chromosome as well as the development of many MHC-linked diseases.