Construction of an expression vector for production of recombinant fluorescent antibodies

Green fluorescent protein can be joined to recombinant antibodies to produce recombinant fluorescent antibodies. Recombinant fluorescent antibodies are functional in different assays in which fluorescent antibodies are traditionally used. Their intrinsic fluorescent renders them directly applicable in many fluorescent-based assays currently used in a wide range of life science. The aim of this study was to manipulate a phagmid antibody vector to produce recombinant fluorescent antibodies by cloning the EGFP gene. With this aim the sequence of a linker peptide and two restriction enzyme sites were added to the EGFP gene and the NcoI restriction site in this gene was removed by means of site-specific mutagenesis. The modified gene was then cloned into pIT2 vector which was subsequently transformed into Escherichia coli. After confirming the sequencing results، this new vector was called pGK-29. Finally، the activities of the recombinant vector were tested by fluorescent microscopy and ELISA experiments. The results revealed that the produced chimeric protein has fluorescent activity and is able to recognize its corresponding antigen.