Abstract：The restriction site-free overlap extension PCR was used to clone two reporter genes with insertion of different yeast introns into the β-galactosidase reporter gene(lacZ).These constructs were subsequently used to examine the effects of different introns on the expression of reporter genes,as well as the efficiency of splicing by β-galactosidase activity assay and real-time(RT) PCR.One major feature of this cloning method is the exemption of introducing additional restriction sites,therefore is particularly important for preservation of some critical sequence context.Comparisons between this method and the In-Fusion cloning technique were addressed in the discussion.
2012, Vol. 36 
