Cloning, expression, and in vitro functional activity assay of phiC31 integrase cDNA in Escherichia coli
Cell Journal [Yakhteh]. 2013; 14 (4): 264-269
em Inglês
| 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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Índice:
IMEMR (Mediterrâneo Oriental)
Assunto principal:
Expressão Gênica
/
Reação em Cadeia da Polimerase
/
DNA Complementar
/
Integrases
/
Clonagem de Organismos
/
DNA Nucleotidiltransferases
/
Eletroforese em Gel de Poliacrilamida
/
Vetores Genéticos
Idioma:
Inglês
Revista:
Cell. J. [Yakhteh]
Ano de publicação:
2013
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