Molecular cloning and characterization of bovine PRKAG3 gene: structure, expression and single nucleotide polymorphism detection.
The protein kinase adenosine monophosphate-activated gamma3-subunit (PRKAG3) gene encodes a muscle-specific isoform of the regulatory gamma-subunit of adenosine monophosphate-activated protein kinase, which plays a key role in regulating energy homeostasis in eucaryotes. It is well known that mutations in the PRKAG3 gene affect high glycogen content in the porcine skeletal muscle and, consequently, meat quality. The genomic structure and sequence of the bovine PRKAG3 were analysed from a Korean cattle BAC clone. The bovine PRKAG3 gene comprises 13 exons and spans approximately 6.8 kb on BTA2. From 5' and 3'-rapid amplification of cDNA ends experiments, the full-length cDNA of bovine PRKAG3 has been identified, encoding a deduced protein of 465 amino acids. Two splice isoforms, generated by the alternative splicing of exon 2, were also identified. Northern blot analysis demonstrated that, similar to other species, the bovine PRKAG3 transcript was only expressed in skeletal muscle. Seven single nucleotide polymorphisms, including two previously identified variants, were detected in four Bos taurus cattle breeds. The bovine PRKAG3 gene described in this study may be involved in muscle-related genetic diseases or meat quality traits in cattle.
Yu
S L
SL
Division of Animal Science and Resources, Research Center for Transgenic Cloned Pigs, Chungnam National University, Daejeon, Korea.
Kim
J E
JE
Chung
H J
HJ
Jung
K C
KC
Lee
Y J
YJ
Yoon
D H
DH
Lee
S H
SH
Choi
I
I
Bottema
C D K
CD
Sang
B C
BC
Lee
J H
JH
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002417:Cattle / Q000235:genetics*; D022202:Chromosomes, Artificial, Bacterial; D003001:Cloning, Molecular; D016000:Cluster Analysis; D019295:Computational Biology; D040461:Gene Components; D015870:Gene Expression; D007723:Korea; D008969:Molecular Sequence Data; D009841:Oligonucleotides; D012150:Polymorphism, Restriction Fragment Length; D011494:Protein Kinases / Q000235:genetics* / Q000378:metabolism; D019105:Random Amplified Polymorphic DNA Technique; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D009841:Oligonucleotides; D011494:Protein Kinases