Cloning and analysis of the cattle FADD gene and construction of eukaryotic expression vector.

【Objective】Cattle FADD gene was cloned and its eukaryotic expression vector was constructed to exploit the gene's regulation role in the procedure of follicular development in bovine ovary.【Method】The FADD gene was amplified in cattle ovary tissue by RT-PCR,the nucleotide sequence in the coding region was analyzed and the termination codon in its cDNA was deleted.The amplified FADD gene was directionally cloned into eukaryotic expression vector pAcGFP-Nl including AcGFP,then identifed by restrictive enzyme BglⅡ/EcoRⅠand sequencing.【Result】The sequence was consistent with that of NCBI,the homology with pig was the highest in nucleotide and amino acid sequence,while that with poultry was the lowest,only 42.5% and 44%.The pAcGFP-N1-bFADD fusion protein recombinant plasmid was successfuly constructed by introducing BglⅡ,EcoRⅠcloning site at two ends of FADD open reading frame and inserting a Kozak sequence before start codon.【Conclusion】Bovine FADD gene was cloned successfully and the pAcGFP-N1-bFADD recombinant plasmid was constructed,it should be helpful for further understanding the mechanism of regulation of FADD on bovine oocyte formation and development.