NTU Undergraduates' research April 2014 - Biosciences | Seite 139

KAM FAI CHEUNG

Generating recombinant DNA to study the trafficking of
histone H2B, PFKFB3 and FZR1 protein in mammalian cell
Abstract
Recombinant DNA is essential for us to study the function of a gene and its ways of ex-pression
in the cell by separating the gene of interest and transferring to another DNA contains
regulatory control sequences. We tried to produce a vector from pSNAPm, us-ing restriction
enzymes of XhoI, ClaI and EcoRI. Sticky ends were made to recombine with the CLIP-tag
sequence from pCLIP-H2B and the sequence encoding FZR1 protein. E. coli played an important
role in the DNA recombination from DNA cloning to the expres-sion of the recombined gene.
The recombination between pSNAPm and pCLIP-H2B as well as pSNAPm and FZR1 were
successful, determined by showing ampicillin re-sistance in E. coli.