title: An optimized yeast cell-free lysate system for in vitro translation of human virus mRNA.
abstract: Yeast (Saccharomyces cerevisiae) as a model organism has long been established to study various aspects of eukaryotic cellular and molecular biology. Cell-free lysates prepared from different yeast strains have been used as a powerful tool to study eukaryotic protein expression in vitro. Recently, we established a yeast cell-free lysate system for in vitro translation long and short L1 capsid gene mRNAs of human papillomavirus type 58. We were able to significantly improve the translation efficiencies of the viral mRNAs in the established system by optimizing the concentrations of potassium and magnesium and controlling the physiological status of the yeast cells used for lysate preparation. We proved that a single specific amino acid can be rate limiting for translation of a target mRNA. Here, we describe the materials, method, and technique used for the development of an efficient yeast cell-free translation system.
authors.affiliation: Department of Pathology, Shandong University School of Medicine, Jinan, People's Republic of China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Wang
authors.forename: Liang
authors.initials: L
authors.lastname: Zhao
authors.forename: Kong-Nan
authors.initials: KN
authors.lastname: Zhao
mesh_terms: D015153:Blotting, Western; D002457:Cell Extracts; D002474:Cell-Free System; D006801:Humans; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000378:metabolism*; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D002457:Cell Extracts; D012333:RNA, Messenger; D012367:RNA, Viral; D014764:Viral Proteins