title: An optimized yeast cell-free system: sufficient for translation of human papillomavirus 58 L1 mRNA and assembly of virus-like particles.
abstract: We developed a yeast cell-free system suitable for in vitro translation of human papillomavirus 58 (HPV58) L1 mRNA. This system was systematically optimized resulting in enhanced translation efficiency. The optimal concentrations of potassium and magnesium ions observed were specific to the HPV58 L1 protein production. Supplementation with sucrose in the preparation of the yeast lysate greatly enhanced its stability. After optimization, protein production in this system was significantly superior to that produced by the rabbit reticulocyte (RRL) system. Finally, we demonstrated for the first time that virus-like particles (VLPs) were assembled from HPV58 L1 capsid protein in the yeast cell-free system. Thus, the system described here is a powerful tool for the HPV L1 protein production and will be useful for the study of VLP assembly and DNA encapsulation.
authors.affiliation: Department of Medical Microbiology, 6504 Building 6, Shandong University School of Medicine, Wenhuaxi Road no. 44, Jinan, Shandong 250012, PR China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Wang
authors.forename: Juan
authors.initials: J
authors.lastname: Liu
authors.forename: Yan
authors.initials: Y
authors.lastname: Zheng
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Yabin
authors.initials: Y
authors.lastname: Zhou
authors.forename: Mei
authors.initials: M
authors.lastname: Qi
authors.forename: Han
authors.initials: H
authors.lastname: Yu
authors.forename: Wei
authors.initials: W
authors.lastname: Tang
authors.forename: Wei Ming
authors.initials: WM
authors.lastname: Zhao
mesh_terms: D036022:Capsid Proteins / Q000235:genetics*; D002474:Cell-Free System; D005818:Genetic Engineering / Q000379:methods*; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D014176:Protein Biosynthesis / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014771:Virion / Q000235:genetics*; D019065:Virus Assembly / Q000235:genetics*
chemical_list: D036022:Capsid Proteins; D009856:Oncogene Proteins, Viral; D012333:RNA, Messenger; D012367:RNA, Viral; C063317:L1 protein, Human papillomavirus type 16