Biological function of basement membrane

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1) We have cloned many overlapping cDNAs coding for the mouse alpha6 (IV) collagen chain and determined primary structure of the alpha6 (IV) chain polypeptide.2) We raised monoclonal antibodies against amino acid sequences from various parts of the mouse alpha6 (IV) collagen polypeptide chain and used them for the tissue distribution. The results indicated that the tissue distribution for the chain was similar to that of the human alpha6 (IV) collagen chain. Interestingly, the distribution of the paired genes ; col4a1/col4a2, col4a3/col4a4, and col4a5/col4a6 is quite synchronous except for the glomerular basement membranes.3) We have some indication that shows alternative splicing in exon 31. Currently, we are characterizing this atypical splicing event in terms of tissue specific expression.4) In order to create a knockout mice for col4a6, we made several constructs including the one where the exon 2 was taken out and the foreign gene, Neo^R gene was inserted instead. The construct was transfected into Stem cells by homologous recombination. Currently, we are characterizing knockout mice to detect any phenotypic changes in heterozygotes and homozygotes.