Abstract
The mRNA differential display technique was performed to investigate the differences of gene expression in the pituitary gland from mini-type Diannan small-ear pigs and large-type Diannan small-ear pigs. One novel gene differentially expressed was identified through semi-quantitative RT-PCR and its cDNA complete sequence was then obtained using the rapid amplification of cDNA ends (RACE) method. Nucleotide sequence of the gene is not homologous to any of the known porcine genes. The sequence analysis revealed that the open reading frame of this gene encodes a protein of 384 amino acids has high homology with the mitogen-activated protein kinase-activated protein kinase 3 (MAPKAPK3) of eight species--cattle (96%), horse (96%), rhesus monkey (93%), rabbit (77%), human (98%), chimpanzee (98%), mouse (94%) and rat (91%)--so that it can be defined as swine MAPKAPK3 gene. This gene was finally assigned GENE ID: 100233192. Computer assisted analysis revealed that the genomic DNA of open reading frame of the swine MAPKAPK3 gene consists of ten exons and nine introns. The phylogenetic tree analysis revealed that the swine MAPKAPK3 gene has a closer genetic relationship with the MAPKAPK3 of cattle. The tissue expression analysis indicated that the MAPKAPK3 mRNA expression levels of large-type Diannan small-ear pigs are ubiquitously increased in various tissues compared to the mini-type Diannan small-ear pigs. Our experiment suggested that the swine MAPKAPK3 gene might play an important role in the growth and development differences between mini-type Diannan small-ear pigs and large-type Diannan small-ear pigs.
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