Authors : YaLin He, Mengliang Liu, XiaoDi Wang, ShanShan Xu and Zhiwen Xu
Abstract: Prokaryotic expression vector of porcine stratifin was constructed to express stratifin in Escherichia coli (E. coli) and eukaryotic expression vector was also constructed. Stratifin cDNA was amplified using the PCR and cloned into pET32a (+) vector to form the recombinant plasmid SFN-pET32a (+); the recombinant plasmid SFN-pET32a (+) was then transformed into prokaryotic expression host E. coli Rosetta using lactose induction, the target protein was purified by using his affinity chromatographic separation. The recombinant protein stratifin was identified by SDS-PAGE. Stratifin cDNA was inserted into pcDNA3.1 (+) to form the recombinant plasmid SFN-pcDNA3.1 (+) and then it was identified by restriction enzyme digestion and DNA sequencing. The recombinant plasmid stratifin was successfully constructed, the recombinant stratifin protein was induced and eukaryotic expression vector was constructed. This study successfully realizes the construction of the prokaryotic and eukaryotic expression vectors and it could be used as a foundation for the future research on the function of this protein.
YaLin He, Mengliang Liu, XiaoDi Wang, ShanShan Xu and Zhiwen Xu, 2012. Construction and Identification of Prokaryotic and Eukaryotic Expression Vectors of Porcine Stratifin. Agricultural Journal, 7: 394-400.