Construction and Fusion Expression of Recombinant IN-1 Single Chain Antibody Expression Vector

Objective: Our aim was to construct and express a recombinant cloning vector bearing the chemically synthesized cDNA of the recombinant IN 1 single chain antibody. Methods: Thirty five segments with the length ranging from 40 to 50 bp were assembled in only one step by using a method of polymerase chain reaction. The entire gene was cloned into pUC18 plasmid. Determined by auto sequencing, the target gene was cloned into prokaryotic expression vector pET28a(+)in fusion form and transformed into E.coli BL21. Results: Sequence analysis showed that the sequence in the gene was almost correct except one base. SDS PAGE analysis showed that a new protein band with molecule weight of 31kd appeared as the expected size. Conclusion: The successful construction and expression of the recombinant vector pET 28a(+) bearing the cDNA might provide material for the further research about the recombinant IN 1 single chain antibody.