Title : ( Cloning, Expression, and In Vitro Functional Activity Assay of phiC31 Integrase cDNA in Escherichia coli )
Authors: Mohammad Hadi Sekhavati , Mojtaba Tahmoorespur , K. Ghaedi , K. Dormiani , Mohammadreza Nassiri , Y. Khazaie , M. Foruzanfar , M. Hosseini , M. H. Nasr Esfahani ,
Full TextAbstract
The aim of present study was cloning and expression of phiC31 integrase cDNA in a bacterial expression vector. Thus, an intra molecular assay vector was applied to show in vitro activity of recombinant protein. Materials and Methods: In this experimental study, phiC31 cDNA was subcloned into a prokaryotic expression vector and transformed into Ecoli Bl21 (DE3). Recombinant phiC31 integrase was purified form the bacterial cell lysates and its activity was verified by an in vitro functional assessment. Results: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the purified phiC31 integrase confirmed the size of protein (70 kDa). Finally, the functionality of purified phiC31 integrase was verified. Conclusion: The results of this study indicated that the purified integrase has a great potential application for in vitro site-specific integration.
Keywords
, phiC31, Site-Specific Integration, E.coli BL21 (DE3)
@article{paperid:1032298,
author = {Sekhavati, Mohammad Hadi and Tahmoorespur, Mojtaba and K. Ghaedi and K. Dormiani and Nassiri, Mohammadreza and Y. Khazaie and M. Foruzanfar and M. Hosseini and M. H. Nasr Esfahani},
title = {Cloning, Expression, and In Vitro Functional Activity Assay of phiC31 Integrase cDNA in Escherichia coli},
journal = {Yakhteh- یاخته},
year = {2013},
volume = {14},
number = {4},
month = {January},
issn = {1561-4921},
pages = {264--269},
numpages = {5},
keywords = {phiC31; Site-Specific Integration; E.coli BL21 (DE3)},
}
%0 Journal Article
%T Cloning, Expression, and In Vitro Functional Activity Assay of phiC31 Integrase cDNA in Escherichia coli
%A Sekhavati, Mohammad Hadi
%A Tahmoorespur, Mojtaba
%A K. Ghaedi
%A K. Dormiani
%A Nassiri, Mohammadreza
%A Y. Khazaie
%A M. Foruzanfar
%A M. Hosseini
%A M. H. Nasr Esfahani
%J Yakhteh- یاخته
%@ 1561-4921
%D 2013
