OBJECTIVETo clone a novel human connexin gene and find out the relationship between this gene and hereditary deafness.

METHODSThrough the basic local alignment search tool (BLAST) analysis against the database of expressed sequence tags (dbEST) of National Center for Biotechnology Information (NCBI) using the coding sequence of mouse Cx 57 gene, 9 novel ESTs were obtained and a contig was assembled. Nested polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE) were performed using primers designed on the contig. A novel gene was obtained and was mapped by homologous analysis against human genome sequence. Mutation analysis was performed in 12 autosomal dominant hereditary deafness families.

RESULTSNine ESTs were obtained by homologous analysis and a contig was assembled. Through nested PCR and RACE, a full length of cDNA was obtained from human liver, kidney Ready cDNA and placenta cDNA library, and was named CX 58. By comparison with human genome sequence, CX 58 was mapped at 1p32.3-p34.1. Mutation analysis of CX 58 was performed in 12 autosomal dominant hereditary deafness families, but no mutation was detected.

CONCLUSIONA novel human connexin gene named CX 58 was cloned and mapped to 1p32.3-p34.1. The mutation of CX 58 may not result in autosomal dominant hereditary deafness.

Animals ; Chromosome Mapping ; Chromosomes, Human, Pair 1 ; genetics ; Cloning, Molecular ; Connexins ; genetics ; DNA, Complementary ; chemistry ; genetics ; Expressed Sequence Tags ; Female ; Humans ; Mice ; Molecular Sequence Data ; Sequence Alignment ; Sequence Analysis, DNA