Development of the Gateway Recycling Cloning System for Multiple Linking of Expression Cassettes in a Defined Order, and Direction on Gateway Compatible Binary Vectors
Tetsuya KimuraDepartment of Sustainable Resource Science, Graduate School of Bioresources, Mie UniversityAkihide NakaoDepartment of Molecular and Functional Genomics, Center for Integrated Research in Science, Shimane UniversitySachiko MURATADepartment of Sustainable Resource Science, Graduate School of Bioresources, Mie UniversityYasuyuki KobayashiDepartment of Sustainable Resource Science, Graduate School of Bioresources, Mie UniversityYuji TanakaDepartment of Molecular and Functional Genomics, Center for Integrated Research in Science, Shimane UniversityKenta ShibaharaDepartment of Molecular and Functional Genomics, Center for Integrated Research in Science, Shimane UniversityTetsu KawazuForestry Research Institute, Oji Paper Co., LtdTsuyoshi NakagawaDepartment of Molecular and Functional Genomics, Center for Integrated Research in Science, Shimane University
ABI
Annotatsiya
We developed the Gateway recycling cloning system, which allows multiple linking of expression cassettes by multiple rounds of the Gateway LR reaction. Employing this system, the recycling donor vector pRED419 was subjected to the first LR reaction with an attR1-attR2 type destination vector. Then conversion vector pCON was subjected to an LR reaction to restore the attR1-attR2 site on the destination vector for the next cloning cycle. By repetition of these two simple steps, we linked four expression cassettes of a reporter gene in Gateway binary vector pGWB1, introduced the constructs into tobacco BY-2 cells, and observed the expression of transgenes.
Mavzular
Identifikatorlar
Iqtiboslar va manbalar
0 ta iqtibos19 ta foydalanilgan manba
Koʻrsatkichlar — AkademScholar · Tez orada