کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
3408542 | 1593511 | 2006 | 6 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Refolding, purification and characterization of replication-initiator protein from soybean-infecting geminivirus Refolding, purification and characterization of replication-initiator protein from soybean-infecting geminivirus](/preview/png/3408542.png)
The replication-initiator protein (Rep) from a soybean-infecting geminivirus was overexpressed in E. coli as a fusion protein with maltose binding protein (MBP). In spite of the presence of the highly soluble MBP as the fusion partner, the overexpressed MBP-Rep fusion protein formed insoluble inclusion bodies. The protein was solubilized from the inclusion bodies and refolded. The refolded MBP-Rep protein was purified using ion exchange and amylose affinity chromatography. The activity of the purified MBP-Rep was assessed using an in vitro cleavage assay. Soluble and stable MBP-Rep protein was obtained in high abundance, providing the feasibility of large-scale production of active Rep protein for functional characterization and X-ray crystallographic structure determination.
Journal: Journal of Virological Methods - Volume 136, Issues 1–2, September 2006, Pages 154–159