Molecular cloning and expression during myogenesis of sequences coding for M-creatine kinase.
Sequences complementary to muscle poly(A)+RNA were cloned in the plasmid pBR322 and the resulting colonies were screened by colony hybridization with labeled cDNA derived from skeletal muscle and smooth muscle (gizzard). The skeletal muscle-specific clones were further screened by RNA blotting hybridization for a muscle mRNA having the size expected for a putative type M creatine kinase (M-CK) mRNA. The remaining clones with the expected hybridization properties were finally characterized by hybrid-selected translation, and a cloned sequence was shown to contain DNA hybridizing to mRNA that could be translated into M-CK. This plasmid, pMCK1, was further characterized by restriction mapping. Blot analysis of total cell RNA from differentiating myogenic cell cultures showed accumulation of M-CK mRNA in cultures older than 42 hr but not in young little-differentiated cultures.
D000818:Animals; D002454:Cell Differentiation; D002642:Chick Embryo; D003001:Cloning, Molecular; D003402:Creatine Kinase / Q000235:genetics*; D004247:DNA / Q000235:genetics; D005786:Gene Expression Regulation; D007527:Isoenzymes / Q000235:genetics; D009130:Muscle, Smooth / Q000502:physiology; D009132:Muscles / Q000166:cytology / Q000502:physiology*
D007527:Isoenzymes; D004247:DNA; D003402:Creatine Kinase