TY - JOUR ID - 250133 TI - Cloning, Expression, and in vitro Functional Activity Assay of phiC31 Integrase cDNA in Escherichia coli JO - Cell Journal (Yakhteh) JA - CELLJ LA - en SN - 2228-5806 AU - Sekhavati, Mohammad Hadi AU - Tahmoorespur, Mojtaba AU - Ghaedi, Kamran AU - Dormiani, Kianoush AU - Nassiri, Mohammad Reza AU - Khazaie, Yahya AU - Foruzanfar, Mahboubeh AU - Hosseini, Morteza AD - . Department of Animal Science, Ferdowsi University of Mashhad, Mashhad, Iran AD - . Department of Molecular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran;. Department of Biology, School of Sciences, University of Isfahan, Isfahan, Iran AD - . Department of Molecular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran;4. Department of Pharmaceutical Biotechnology, Faculty of Pharmacy and Pharmaceutical Scien AD - . Department of Molecular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran AD - 5. Department of Reproductive Biotechnology at Reproductive Biomedicine Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran Y1 - 2013 PY - 2013 VL - 14 IS - 4 SP - 264 EP - 269 KW - phiC31 KW - Site KW - Specific Integration KW - BL21 (DE3) DO - N2 - Objective 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 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. Results: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the puriļ¬ed 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. UR - https://www.celljournal.org/article_250133.html L1 - https://www.celljournal.org/article_250133_e1369b51494bcfee9a5fcab36febf244.pdf ER -