title: A Rapid and Efficient Method for Construction of an Infectious Clone of Tomato yellow leaf curl virus.
abstract: Tomato yellow leaf curl virus (TYLCV), a member of the genus Begomovirus, is responsible for one of the most devastating viral diseases in tomato-growing countries and is becoming a serious problem in many subtropical and tropical countries. The climate in Korea is getting warmer and developing subtropical features in response to global warming. These changes are being accompanied by TYLCV, which is now becoming a large problem in the Korean tomato industry. The most effective way to reduce damage caused by TYLCV is to breed resistant varieties of tomatoes. To accomplish this, it is necessary to establish a simple inoculation technique for the efficient evaluation of resistance to TYLCV. Here, we present the rolling circle amplification (RCA) method, which employs a bacteriophage using phi-29 DNA polymerase for construction of infectious TYLCV clones. The RCA method is simple, does not require sequence information for cloning, and is less expensive and time consuming than conventional PCR based-methods. Furthermore, RCA-based construction of an infectious clone can be very useful to other emerging and unknown geminiviruses in Korea.
authors.affiliation: Department of Microbiology, Pusan National University, Busan 609-735, Korea.
authors.forename: Bongjun
authors.initials: B
authors.lastname: Bang
authors.affiliation: Department of Microbiology, Pusan National University, Busan 609-735, Korea.
authors.forename: Jongyun
authors.initials: J
authors.lastname: Lee
authors.affiliation: Department of Microbiology, Pusan National University, Busan 609-735, Korea.
authors.forename: Sunyoung
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Microbiology, Pusan National University, Busan 609-735, Korea.
authors.forename: Jungwook
authors.initials: J
authors.lastname: Park
authors.affiliation: Department of Microbiology, Pusan National University, Busan 609-735, Korea.
authors.forename: Thao Thi
authors.initials: TT
authors.lastname: Nguyen
authors.affiliation: Department of Microbiology, Pusan National University, Busan 609-735, Korea.
authors.forename: Young-Su
authors.initials: YS
authors.lastname: Seo
keywords: Tomato yellow leaf curl virus; infectious clone; rolling circle amplification; tomato