An optimized yeast cell-free system: sufficient for translation of human papillomavirus 58 L1 mRNA and assembly of virus-like particles.
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.
Wang
Xiao
X
Department of Medical Microbiology, 6504 Building 6, Shandong University School of Medicine, Wenhuaxi Road no. 44, Jinan, Shandong 250012, PR China.
Liu
Juan
J
Zheng
Yan
Y
Li
Jing
J
Wang
Hong
H
Zhou
Yabin
Y
Qi
Mei
M
Yu
Han
H
Tang
Wei
W
Zhao
Wei Ming
WM
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*
D036022:Capsid Proteins; D009856:Oncogene Proteins, Viral; D012333:RNA, Messenger; D012367:RNA, Viral; C063317:L1 protein, Human papillomavirus type 16