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A High Yield and Cost-efficient Expression System of Human Granzymes in Mammalian Cells
Published on: June 10, 2015
[Construction and expression of recombinant antibody/granzyme B containing truncated translocating peptide]
Aim:
To construct an eukaryotic expression vector for recombinant antibody/granzyme B gene containing truncated translocating peptide and express it in HER-2(+) SKBR-3 cells and HER-2(-) Hela cells.
Methods:
PCR amplication was used to obtain recombinant DNA encoding antibody/granzyme B containing truncated translocating peptide DNA, and then the DNA fragment was cloned into eukaryotic expression vector pCMV-e23sFv-PE40. After being transfected into SKBR-3 cells and Hela cells, the expression of target gene and its effect on cellular morphology were detected by immunocytochemical staining.
Results:
The eukaryotic expression vector encoding recombinant antibody/granzyme B containing truncated translocating peptide was successfully constructed. e23sFv-FSD-GrB protein was expressed in most of Hela cells and had no effect on Hela cells, but the transfected SKBR-3 cells which also express e23sFv-FSD-GrB protein exhibited condensed nucleus and cytoplasm.
Conclusion:
The construction of eukaryotic expression vector of recombinant antibody/granzyme B gene containing truncated translocating peptide lays the foundation for further determination of minimal fragment responsible for translocation.
