Cloning, Expression, and In Vitro Functional Activity Assay of phiC31 Integrase cDNA in Escherichia coli
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 in...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Royan Institute (ACECR), Tehran
2013-01-01
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| Series: | Cell Journal |
| Subjects: | |
| Online Access: | http://celljournal.org/library/upload/article/af_33756277423752622223626756247246557883623-Sekhavati-1.pdf |
| _version_ | 1818607021438533632 |
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| author | Mohammad Hadi Sekhavati Mojtaba Tahmoorespur Kamran Ghaedi Kianoush Dormiani, Pharm Mohammad Reza Nassiri Yahya Khazaie Mahboubeh Foruzanfar Morteza Hosseini Mohammad Hossein Nasr Esfahani |
| author_facet | Mohammad Hadi Sekhavati Mojtaba Tahmoorespur Kamran Ghaedi Kianoush Dormiani, Pharm Mohammad Reza Nassiri Yahya Khazaie Mahboubeh Foruzanfar Morteza Hosseini Mohammad Hossein Nasr Esfahani |
| author_sort | Mohammad Hadi Sekhavati |
| collection | DOAJ |
| description | 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 purifi 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. |
| first_indexed | 2024-12-16T14:20:07Z |
| format | Article |
| id | doaj.art-f413d7e9231247edb013092fa5c9ebfc |
| institution | Directory Open Access Journal |
| issn | 2228-5806 2228-5814 |
| language | English |
| last_indexed | 2024-12-16T14:20:07Z |
| publishDate | 2013-01-01 |
| publisher | Royan Institute (ACECR), Tehran |
| record_format | Article |
| series | Cell Journal |
| spelling | doaj.art-f413d7e9231247edb013092fa5c9ebfc2022-12-21T22:28:32ZengRoyan Institute (ACECR), TehranCell Journal2228-58062228-58142013-01-01144264269Cloning, Expression, and In Vitro Functional Activity Assay of phiC31 Integrase cDNA in Escherichia coliMohammad Hadi SekhavatiMojtaba TahmoorespurKamran GhaediKianoush Dormiani, PharmMohammad Reza NassiriYahya KhazaieMahboubeh ForuzanfarMorteza HosseiniMohammad Hossein Nasr EsfahaniObjective: 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 purifi 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.http://celljournal.org/library/upload/article/af_33756277423752622223626756247246557883623-Sekhavati-1.pdfphiC31Site-Specific IntegrationE.coli BL21 (DE3) |
| spellingShingle | Mohammad Hadi Sekhavati Mojtaba Tahmoorespur Kamran Ghaedi Kianoush Dormiani, Pharm Mohammad Reza Nassiri Yahya Khazaie Mahboubeh Foruzanfar Morteza Hosseini Mohammad Hossein Nasr Esfahani Cloning, Expression, and In Vitro Functional Activity Assay of phiC31 Integrase cDNA in Escherichia coli Cell Journal phiC31 Site-Specific Integration E.coli BL21 (DE3) |
| title | Cloning, Expression, and In Vitro Functional Activity Assay of phiC31 Integrase cDNA in Escherichia coli |
| title_full | Cloning, Expression, and In Vitro Functional Activity Assay of phiC31 Integrase cDNA in Escherichia coli |
| title_fullStr | Cloning, Expression, and In Vitro Functional Activity Assay of phiC31 Integrase cDNA in Escherichia coli |
| title_full_unstemmed | Cloning, Expression, and In Vitro Functional Activity Assay of phiC31 Integrase cDNA in Escherichia coli |
| title_short | Cloning, Expression, and In Vitro Functional Activity Assay of phiC31 Integrase cDNA in Escherichia coli |
| title_sort | cloning expression and in vitro functional activity assay of phic31 integrase cdna in escherichia coli |
| topic | phiC31 Site-Specific Integration E.coli BL21 (DE3) |
| url | http://celljournal.org/library/upload/article/af_33756277423752622223626756247246557883623-Sekhavati-1.pdf |
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