Cloning, expression, and in vitro functional activity assay of phiC31 integrase cDNA in Escherichia coli
Cell Journal [Yakhteh]. 2013; 14 (4): 264-269
in English
| IMEMR
| ID: emr-140460
ABSTRACT
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. In this experimental study, phiC31 cDNA was subcloned into a prokaryotic expression vector and transformed into E.coli Bl21 [DE3]. Recombinant phiC31 integrase was purified form the bacterial cell lysates and its activity was verified by an in vitro functional assessment. 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. The results of this study indicated that the purified integrase has a great potential application for in vitro site-specific integration
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Index:
IMEMR (Eastern Mediterranean)
Main subject:
Gene Expression
/
Polymerase Chain Reaction
/
DNA, Complementary
/
Integrases
/
Cloning, Organism
/
DNA Nucleotidyltransferases
/
Electrophoresis, Polyacrylamide Gel
/
Genetic Vectors
Language:
English
Journal:
Cell. J. [Yakhteh]
Year:
2013
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