کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
3406644 | 1223581 | 2013 | 8 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Expression in tobacco and purification of beak and feather disease virus capsid protein fused to elastin-like polypeptides Expression in tobacco and purification of beak and feather disease virus capsid protein fused to elastin-like polypeptides](/preview/png/3406644.png)
• BFDV capsid protein (CP) and truncated CP (ΔN40 CP) were transiently expressed in tobacco.
• Fusion of CP or ΔN40 CP to elastin-like polypeptides (ELPs) increased yields.
• The purification of a 26 kDa ELP by inverse transition cycling (ITC) was optimised.
• Membrane filtration ITC gave the highest recovery of ΔN40 CP-ELP fusion protein.
Psittacine beak and feather disease, caused by beak and feather disease virus (BFDV), is a threat to endangered psittacine species. There is currently no vaccine against BFDV, which necessitates the development of safe and affordable vaccine candidates. A subunit vaccine based on BFDV capsid protein (CP), the major antigenic determinant, expressed in the inexpensive and highly scalable plant expression system could satisfy these requirements. Full-length CP and a truncated CP (ΔN40 CP) were transiently expressed in tobacco (Nicotiana benthamiana) as fusions to elastin-like polypeptide (ELP). These two proteins were fused to ELPs of different lengths in order to increase expression levels and to provide a simple means of purification. The ELP fusion proteins were purified by inverse transition cycling (ITC) and it was found that a membrane filtration-based ITC method improved the recovery of ΔN40 CP-ELP51 fusion protein relative to a centrifugation-based method.
Journal: Journal of Virological Methods - Volume 191, Issue 1, July 2013, Pages 55–62