PARTIAL TITIN CDNA SEQUENCE ISOLATED FROM RABBIT CARDIAC-MUSCLE RNA

Two regions of the rabbit cardiac titin cDNA were amplified from rabbit cardiac muscle total RNA using primers based on rabbit skeletal muscle titin (connectin) cDNAs. These 1.7 kb and 1.5 kb RNA-PCR products were based on the 3' regions of the skeletal muscle titin clones CE12 and MS2, respectively. The cDNA sequence of the 1.7 kb product was extended an additional 1.5 kb by a novel 3' extension technique which used random primers in RNA-PCR. The cardiac titin cDNAs were 99% identical in nucleotide sequence to their skeletal muscle counterparts and predicted two types of 100-residue repeats, Southern blot analysis suggested that both cardiac and skeletal titin are encoded by the same gene. PCR amplification of human genomic DNA with titin specific primers indicated that there is strong sequence similarity between rabbit and human titin sequences. The successful amplification of a 907 basepair region from human genomic DNA suggested that titin contains large exons which span multiple motif borders. This may be particularly advantageous in the processing of such a large RNA transcript.