A universal mini-vector and an annealing of PCR products (APP)-based cloning strategy for convenient molecular biological manipulations
| Autor: | Yifei Liu, Darren J. Hart, Shumin Xu, Hongling Wang, Yifeng Zhang, Tuoping Li, Xia Liu, Yingfeng An, Gao Herui, Song Gao |
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| Přispěvatelé: | College of Biosciences and Biotechnology, Shenyang Agricultural University, College of Food Science, Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA) |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
T-vector Computer science Genetic Vectors Biophysics Computational biology Molecular cloning Polymerase Chain Reaction Biochemistry TA cloning 03 medical and health sciences Plasmid MESH: Genetic Vectors MESH: Plasmids Ligation-independent cloning Methods Multiple cloning site False positive paradox MESH: Cloning Molecular Cloning Molecular MESH: Methods Ligation Molecular Biology Sticky-end ligation 2. Zero hunger [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry Molecular Biology/Structural Biology [q-bio.BM] MESH: Polymerase Chain Reaction Cell Biology MESH: Ligation 030104 developmental biology Protein expression Restriction digest Plasmids |
| Zdroj: | Biochemical and Biophysical Research Communications Biochemical and Biophysical Research Communications, Elsevier, 2018, 497 (4), pp.978-982. ⟨10.1016/j.bbrc.2018.02.099⟩ Biochemical and Biophysical Research Communications, 2018, 497 (4), pp.978-982. ⟨10.1016/j.bbrc.2018.02.099⟩ |
| ISSN: | 0006-291X 1090-2104 |
| DOI: | 10.1016/j.bbrc.2018.02.099 |
| Popis: | International audience; Currently, the most widely used strategies for molecular cloning are sticky-end ligation-based cloning, TA cloning, blunt-end ligation-based cloning and ligase-independent cloning. In this study we have developed a novel mini-vector pANY1 which can simultaneously meet the requirements of all these cloning strategies. In addition, the selection of appropriate restriction digestion sites is difficult in some cases because of the presence of internal sites. In this study, an annealing of PCR products (APP)-based sticky-end cloning strategy was introduced to avoid this issue. Additionally, false positives occur during molecular cloning, which increases the workload of isolating positive clones. The plasmid pANY1 contains a ccdB cassette between multiple cloning sites, which efficiently avoids these false positives. Therefore, this mini-vector should serve as a useful tool with wide applications in biosciences, agriculture, food technologies, etc. |
| Databáze: | OpenAIRE |
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