Construction and Fusion Expression of Expression Vector of Recombinant Anti-NI-35 Single Chain Antibody

Objective: Our aim was to construct and express a recombinant cloning vector bearing the chemically synthesized cDNA of the recombinant anti-NI-35 single-chain antibody. Methods: Thirty-five segments of 40 to 50 bp were assembled in only one step by a PCR approach. The entire gene was cloned into pUC18 plasmid. After determined by auto-sequencing, the target gene was cloned into prokaryotic expression vector pET28a(+)in fusion form and transformed into. E.coli BL21. Result: Sequence analysis showed that the splicing order, the direction and the sequence in the gene were correct. SDS-PAGE analysis showed a new protein band with molecule weight of 31kDa, which appeared as expected. Conclusion: The successful construction and expression of the recombinant vector pET-28a(+) bearing the cDNA might provide basis for further research about the anti-NI-35 single-chain antibody.