[Construction of a fused fpg gene by using TP-PCR method].
TP-PCR,a method developed for fusion gene construction without the use of endonuclease and ligase, was performed to construct a fused fpg gene. The TP-PCR reaction system contained three primers and two templates and resulting PCR product, fused fpg gene, consisted of three sections: pheB gene, which was responsible for catechol 2,3-dioxygenase, gfp gene for GFP protein and the intermediate ligation segment which was designed for the correct expression of the fusion gene. The result in this paper showed that the TP-PCR method is one of rapid and convenient methods for fused gene construction.
Liu
He
H
Department of Environmental Engineering, Zhejiang Uinversity, Hangzhou 310 029, China. liuhe@zju.edu.cn
Chen
Ying-Xu
YX
Zhang
Wen-Bo
WB
Jin
Yong-Feng
YF
D050557:Catechol 2,3-Dioxygenase / Q000235:genetics*; D003001:Cloning, Molecular; D017931:DNA Primers; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D017422:Sequence Analysis, DNA; D013698:Templates, Genetic
D017931:DNA Primers; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D050557:Catechol 2,3-Dioxygenase