Cloning, nucleotide sequence and characterization of a full-length cDNA encoding the myosin heavy chain from adult chicken pectoralis major muscle

Gene. 1997 Oct 15;199(1-2):265-70. doi: 10.1016/s0378-1119(97)00386-7.

Abstract

Four cDNA clones, encoding the chicken adult sarcomeric MyHC, have been isolated from a pectoralis major muscle cDNA library using gene-specific DNA probes. These clones were sequenced and then subcloned into a full-length, 6-kb, chicken adult sarcomeric MyHC cDNA. The entire cDNA consists of 5873 nucleotides with 19 bp 5'-untranslated region and 34 bp 3'-untranslated region. The complete cDNA encodes a 1939-aa polypeptide whose molecular weight is 223 kDa. The calculated isoelectric point of this protein is approximately 5.7. Analysis of the deduced amino acid sequence and comparison with a previously published amino-acid sequence of the same MyHC isoform reveals that six amino acid residues are different. Hydrophilicity analysis of this adult MyHC amino-acid sequence shows a similar pattern as the embryonic MyHC. A recombinant baculovirus, carrying this full-length adult MyHC cDNA, has also been generated and expressed in the Sf9 insect cell line. A approximately 220-kDa recombinant MyHC was synthesized and reacted specifically with chicken adult MyHC monoclonal antibodies.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Amino Acids / analysis
  • Animals
  • Antibodies, Monoclonal
  • Baculoviridae / genetics
  • Base Sequence
  • Cell Line
  • Chickens
  • Cloning, Molecular
  • DNA, Complementary / genetics*
  • Genetic Vectors / genetics
  • Molecular Sequence Data
  • Molecular Weight
  • Myosin Heavy Chains / analysis
  • Myosin Heavy Chains / chemistry
  • Myosin Heavy Chains / genetics*
  • Pectoralis Muscles / physiology*
  • Recombinant Fusion Proteins
  • Sarcomeres / genetics
  • Sequence Analysis, DNA
  • Spodoptera

Substances

  • Amino Acids
  • Antibodies, Monoclonal
  • DNA, Complementary
  • Recombinant Fusion Proteins
  • Myosin Heavy Chains

Associated data

  • GENBANK/U87231