Expression of Chicken Neurexophilin 1 Gene in Pichia pastoris

doi:10.13560/j.cnki.biotech.bull.1985.2015.04.030

Abstract

Abstract: Neurexophilin 1 is the differentially expressed gene at the chicken utero-vaginal junction. According to the mRNA sequence of chicken nxph1 and codon bias of Pichia pastoris, the modified DNA sequence for chicken nxph1 which encodes the same amino acid sequence as that of chicken nxph1were synthesized. Specific primers were designed to amplify part of the synthesized DNA sequence whose expression product has no signal peptide. The two sequences were inserted into pPICZαA vector to construct the recombinant pPICZαA/nxph1 and pPICZαA/△nxph1(without signal peptide), and then the recombinant plasmids were transfected into Pichia pastoris GS115 by electroporation. The positive recombinant strains were inducted to express protein by addition of 1 % methanol. The expression protein was displayed on the Tricine-SDS- PAGE electrophoresis and Western blot. The result showed that chicken NXPH1 protein and △NXPH1 protein were successfully expressed, and the molecular weight of the protein was approximately 29 kD, which provided foundations for analyzing functions of NXPH1 in chicken reproductive tract.

Key words: nxph 1, Pichia pastoris, gene expression

Chen Meiling, Nie Bin, Jiang Xunping, Liu Guiqiong. Expression of Chicken Neurexophilin 1 Gene in Pichia pastoris[J]. Biotechnology Bulletin, 2015, 31(3): 213-217.

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