Molecular Cloning of WAS Protein Family, Member 1 (WAVE1) in Porcine Oocytes

Oocytes have ability to remodel epigenetic marks of somatic cells therefore reprogram the somatic cells into embryonic state. Unique reprogramming factors which reside in oocytes are thought to be contributing to this reprogramming process. However, specific oocytes factors are not well-characterized. The WAVE1 gene, derived from Xenopus eggs, is shown to reprogram murine somatic cells into pluripotent state. The gene is specifically related to transcriptional activation of embryonic genes by affecting RNA polymerase II activity. When amino acid sequence of WAVE1 was compared among various species, it was highly conserved although little similarity was found at the nucleotide level. Because the gene is highly conserved, we hypothesized that the gene has a similar role in pigs as a reprogramming factor. To clone the full coding sequence of porcine WAVE1, we identified a porcine homolog of WAVE1 from GenBank. RT-PCR results using primers designed based on the sequence demonstrate that the WAVE1 is expressed in porcine oocytes. To clone the entire coding sequence of porcine WAVE1, predicted sequence of porcine WAVE1 and ESTs extending sequences of the predicted WAVE1 were identified from GenBank. Primers designed based on the sequence information could amplify the entire coding sequence of porcine WAVE1 from cDNA, derived from oocytes. Sequencing results indicate that the porcine WAVE1 has high identity with human and Xenopus at the amino acid level; 99% and 82% respectively. The coding sequence of porcine WAVE1 will be overexpressed in somatic cells to examine its ability to reprogram somatic cells in pigs.