Abstract
Histidine-tagged proteins have been purified with immobilized nickel affinity chromatography. This method, however, bears some serious disadvantages, including carcinogenicity and potential leakage of the nickel and the handling and disposal costs. We developed a novel purification process for water-soluble histidine-tagged green fluorescent protein from the cocoons of transgenic silkworms, unlike conventional E. coli expression system which sometimes produced insoluble and inactive inclusion bodies. 60.3% extraction efficiency was achieved with 50mM Tris containing 150mM NaCl, pH 7.5. The extract was purified in one step on zinc-immobilized hydroxyapatite eluted with phosphate buffer. Purity was 91.2% with a recovery of 30.0μg from 160mg of silk fiber. This purification method may provide a simple non-toxic alternative to nickel-immobilized metal affinity chromatography for purification of other histidine-tagged proteins.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
