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2012 | 30 | 3 |
Tytuł artykułu

cDNA cloning of porcine CDH1 and its expression profile in porcine early parthenotes*

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The study aimed at cloning the complete cDNA sequence of porcine CDH1 gene and detecting its expression in different developmental stages of early parthenotes. After cloning the CDH1 gene from porcine oviduct using rapid amplification of cDNA ends (RACE) method, the sequence analysis revealed that porcine CDH1 gene complete cDNA nucleotide sequence amounted to 4283 bp including 2652 bp of open reading frame (ORF), 105 bp of 5’ untranslated region (UTR) and 1526 bp of 3’ UTR. The ORF encoded a deduced protein precursor of 97 kD with 883 amino acid residues. The precursor protein including signal peptide, extracellular region, membrane-spanning region and cytoplasmic region had a single transmembrane structure, and its extracellular region had HAV motif and some Ca2+ binding regions. The porcine CDH1 protein showed high homology with cattle (89%), horse (87%), dog (86%), human (84%), chimpanzee (83%) and mouse (83%) CDH1. The results from RT-PCR and real-time PCR indicated that CDH1 gene could be Expressem in both immature and mature oocytes, as well as early parthenotes (2-, 4-, 8-cell embryos, morula, blastocysts). The data of real-time PCR showed that the expression of CDH1 was the highest at 2-cell parthenogenetic embryos stage, and then it decreased significantly till 8-cell embryos stage; the second increased expression level occurred at morulas stage, and then also showed a diminishing trend in the following developmental stage. This is the first report of cloning and analysing the porcine CDH1 cDNA that provides critical information for further research of its functions in pig embryo development.
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