title: Optimal transfection methods and comparison of PK-15 and Dulac cells for rescue of chimeric porcine circovirus type 1-2.
abstract: A chimeric porcine circovirus type 1-2 (PCV1-2) infectious DNA clone has low transfection efficiency and exhibits low levels of proliferation. Electroporation and lipofection parameters were optimized for PK-15 and Dulac cells with the purpose of increasing the efficiency for rescuing infectious PCV1-2. Titers of PCV1-2 in Dulac cells were 100-fold higher than those in PK-15 cells following transfection. The electroporation efficiency into Dulac cells was high when three 400 μs pulses at 250 V with 6 μg of plasmid DNA was used, lipofection efficiency was high when the ratio of DNA to transfection reagent was 1:3. The proportion of infected cells was 55.6% compared with 44.2%, for the electroporation and lipofection techniques respectively. Virus titers were higher in Dulac cells, from 10(4.44) to 10(5.32)TCID50/mL compared with 10(1.90)-10(3.38)TCID(50)/mL for PK-15 cells. Dulac cells were more permissive to PCV1-2 than PK-15 cells regardless of the transfection technique.
authors.affiliation: Animal Infectious Disease Laboratory, Ministry of Agriculture, Jiangsu Co-Innovation Centre for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu 225009, China.
authors.forename: Jizong
authors.initials: J
authors.lastname: Li
authors.affiliation: Animal Infectious Disease Laboratory, Ministry of Agriculture, Jiangsu Co-Innovation Centre for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu 225009, China.
authors.forename: Tianqi
authors.initials: T
authors.lastname: Yu
authors.affiliation: Animal Infectious Disease Laboratory, Ministry of Agriculture, Jiangsu Co-Innovation Centre for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu 225009, China.
authors.forename: Jinzhu
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Animal Infectious Disease Laboratory, Ministry of Agriculture, Jiangsu Co-Innovation Centre for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu 225009, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Tu
authors.affiliation: Animal Infectious Disease Laboratory, Ministry of Agriculture, Jiangsu Co-Innovation Centre for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu 225009, China. Electronic address: gsong@yzu.edu.cn.
authors.forename: Song
authors.initials: S
authors.lastname: Gao
authors.affiliation: Animal Infectious Disease Laboratory, Ministry of Agriculture, Jiangsu Co-Innovation Centre for the Prevention and Control of Important Animal Infectious Disease and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu 225009, China.
authors.forename: Xiufan
authors.initials: X
authors.lastname: Liu
mesh_terms: D000818:Animals; D002460:Cell Line; D017925:Circovirus / Q000235:genetics* / Q000254:growth & development*; D018274:Electroporation / Q000379:methods; D011995:Recombination, Genetic; D014162:Transfection / Q000379:methods*; D019562:Viral Load; D014773:Virology / Q000379:methods
keywords: Chimeric porcine circovirus type 1-2; Dulac cells; Electroporation; Lipofection; PK-15 cells