TY  - BOOK
AU  - Blawid,R.
AU  - Nagata,T.
TI  - Construction of an infectious clone of a plant RNA virus in a binary vector using one-step Gibson Assembly
KW  - FULL-LENGTH CDNA CLONE
KW  - GIBSON ASSEMBLY
KW  - SEAMLESS CLONING
KW  - TYMOVIRUS
N2  - The construction of full-length infectious clones of RNA viruses is often laborious due to the many cloning steps required and the DNA exclusion within the plasmid during Escherichia coli transformation. We demonstrate single-step cloning procedure of an infectious cDNA of the tomato blistering mosaic virus (ToBMV)using Gibson Assembly (GA), which drastically reduces the number of cloning steps. By agro-inoculation with the construct obtained by this procedure, ToBMV was recovered six days post-inoculation in Nicotiana benthamiana plants. The symptoms induced by the recovered virus were indistinguishable from those caused by the wild-type virus. We conclude that the GA is very useful method particularly to construct a full-length cDNA clone of a plant RNA virus in a binary vector
UR  - https://drive.google.com/file/d/1q8wEqKxx0QD65ZOfX60ve6WaKJzYOgYW/view?usp=drivesdk
ER  -