Molecular characterization, expression patterns, and polymorphism of a differentially expressed porcine gene (PYGM) isolated by suppression subtractive hybridization and two-dimensional gel electrophoresis analysis.
Suppression subtractive hybridization was performed to detect the differences in gene expression of porcine longissimus dorsi muscles between Large White and Chinese Meishan pigs. An upregulated gene in Large White that shared high homology with human muscle glycogen phosphorylase (PYGM) was identified. The porcine PYGM gene contains an open reading frame encoding 842 amino acid residues with 26 and 283 nucleotides in the 5' and 3' untranslated regions, respectively. Tissue distribution analysis indicated that porcine PYGM mRNAs are highly expressed in all tissues. Expression pattern of PYGM was similar in the two breeds. Both breeds had the highest expression levels when 120 days old (p<0.01), and PYGM was upregulated during skeletal muscle development. A similar expression pattern of PYGM in protein level was also observed by differential proteome analysis of skeletal muscle development using two-dimensional gel electrophoresis and mass spectroscopy. The mRNA abundance of PYGM in Large White was higher than Meishan at all four stages (p<0.05). Moreover, a G/T mutation in exon 8 was identified and association analysis with meat quality traits showed that it was significantly associated with lean meat percentage (p<0.05). Our data may provide further insight into the molecular mechanisms responsible for breed-specific differences in porcine growth and meat quality.
Institute of Genetics, College of Life Sciences, Xinyang Normal University, Xinyang, Henan, P.R. China.
Yongjie
Y
Xu
Wenmin
W
Yu
Xiaoting
X
Feng
Hongtao
H
Xie
Yuanzhu
Y
Xiong
D000818:Animals; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D005787:Gene Frequency; D005838:Genotype; D024982:Glycogen Phosphorylase, Muscle Form / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D008460:Meat / Q000592:standards; D024510:Muscle Development / Q000235:genetics; D018482:Muscle, Skeletal / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010802:Phylogeny; D017354:Point Mutation; D020641:Polymorphism, Single Nucleotide; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013045:Species Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013552:Swine / Q000145:classification / Q000235:genetics*; D013997:Time Factors
D018076:DNA, Complementary; D024982:Glycogen Phosphorylase, Muscle Form