Molecular cloning and expression during myogenesis of sequences coding for M-creatine kinase.
1982; National Academy of Sciences; Volume: 79; Issue: 21 Linguagem: Inglês
10.1073/pnas.79.21.6589
ISSN1091-6490
AutoresUrs B. Rosenberg, G Kunz, A Frischauf, Hans Lehrach, R. Mähr, Hans M. Eppenberger, Jean‐Claude Perriard,
Tópico(s)Muscle Physiology and Disorders
ResumoSequences 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.
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