CLONING, NUCLEOTIDE-SEQUENCE AND CHARACTERIZATION OF A FULL-LENGTH CDNA-ENCODING THE MYOSIN HEAVY-CHAIN FROM ADULT CHICKEN PECTORALIS MAJORMUSCLE

Four cDNA clones, encoding the chicken adult sarcomeric MyHC, have bee n isolated from a pectoralis major muscle cDNA library using gene-spec ific DNA probes. These clones were sequenced and then subcloned into a full-length, 6-kb, chicken adult sarcomeric MyHC cDNA. The entire cDN A consists of 5873 nucleotides with 19 bp 5'-untranslated region and 3 4 bp 3'-untranslated region. The complete cDNA encodes a 1939-aa polyp eptide whose molecular weight is similar to 223 kDa. The calculated is oelectric point of this protein is similar to 5.7. Analysis of the ded uced amino acid sequence and comparison with a previously published am ino-acid sequence of the same MyHC isoform reveals that six amino acid residues are different. Hydrophilicity analysis of this adult MyHC am ino-acid sequence shows a similar pattern as the embryonic MyHC. A rec ombinant baculovirus, carrying this full-length adult MyHC cDNA, has a lso been generated and expressed in the Sf9 insect cell line. A simila r to 220-kDa recombinant MyHC was synthesized and reacted specifically with chicken adult MyHC monoclonal antibodies. (C) 1997 Elsevier Scie nce B.V.