title: Cloning the partial cDNAs of mu-calpain and m-calpain from porcine skeletal muscle.
abstract: Calpains are non-lysosomal proteases involved in myofibrillar protein degradation. To facilitate studying the expression of the porcine calpain genes and their influence on protein accretion, we have cloned partial cDNAs for mu- and m-calpain from porcine skeletal muscle via PCR amplification. A 289 bp fragment for mu-calpain and a 629 bp fragment for m-calpain were cloned into the EcoRV site of pBluescript II KS+ vector. The nucleotide sequence for porcine mu-calpain and m-calpain were 92% and 90% identical to corresponding regions of rabbit mu- and m-calpain, respectively. The deduced amino acid sequences for both mu- and m-calpain share 94% identity with respective rabbit mu- and m-calpains. Isoform specificity was validated by Southern hybridization of mu- and m-calpain probes with cloned mu- and m-calpain fragments and Northern hybridization with pig muscle mRNA. These clones will be used to evaluate the role of calpain expression in muscle hypertrophy.
authors.affiliation: Department of Animal Sciences, Purdue University, West Lafayette, Indiana 47907.
authors.forename: W
authors.initials: W
authors.lastname: Sun
authors.forename: S Q
authors.initials: SQ
authors.lastname: Ji
authors.forename: P J
authors.initials: PJ
authors.lastname: Ebert
authors.forename: C A
authors.initials: CA
authors.lastname: Bidwell
authors.forename: D L
authors.initials: DL
authors.lastname: Hancock
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002154:Calpain / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D015870:Gene Expression; D008969:Molecular Sequence Data; D009132:Muscles / Q000737:chemistry*; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D011817:Rabbits; D016415:Sequence Alignment; D013552:Swine
chemical_list: D018076:DNA, Complementary; D002154:Calpain