Recombinant Expressed Vector pET32a (+) S Constructed by Ligation Independent Cloning
AbstractThe aim of this work was to develop a new method for constructing vectors, named ligation-independent cloning (LIC) method. We constructed the S label expression vector and recombinant pET32a (+) S-phoN2 by LIC. The recombinant proteins were expressed in E. coli at a high level, and then the specificity of the recombinant proteins was identified by western blot. The target band was detected by S monoclonal antibody and Apyrase polyclonal antibodies but not Trx monoclonal antibody and HIS monoclonal antibody. Finally, we obtained protein Apyrase in E. coli (BL21), with a protein-only expression S tag. Collectively, our results demonstrated that LIC is effective for the construction of new vectors and recombinant plasmids. Free from the limitations of restriction enzyme sites and with a higher positive rate, LIC processes should find broad applications in molecular biology research. View Full-Text
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Wang, Y.; Gong, G.-H.; Wei, C.-X.; Liang, L.; Zhang, B. Recombinant Expressed Vector pET32a (+) S Constructed by Ligation Independent Cloning. Molecules 2014, 19, 16179-16189.
Wang Y, Gong G-H, Wei C-X, Liang L, Zhang B. Recombinant Expressed Vector pET32a (+) S Constructed by Ligation Independent Cloning. Molecules. 2014; 19(10):16179-16189.Chicago/Turabian Style
Wang, Yu; Gong, Guo-Hua; Wei, Cheng-Xi; Liang, Long; Zhang, Bin. 2014. "Recombinant Expressed Vector pET32a (+) S Constructed by Ligation Independent Cloning." Molecules 19, no. 10: 16179-16189.