Hi all, If anybody is interested in non-viral stable expression, we have a piggybac-based, doxycycline-inducible system. It is the reference 40 in the lentiviral paper that Tomas directed to. We’d be happy to distribute the plasmids.
Zhijie > On Jan 25, 2020, at 3:26 AM, Tomas Malinauskas <tomas.malinaus...@gmail.com> > wrote: > > Hi Gloria, > > two key papers describing expression (transient and lentivirus-based) > of proteins in HEK293 cells we use to make milligrams of proteins for > crystallization and cryo-EM: > > Acta Crystallogr D Biol Crystallogr. 2006 Oct;62(Pt 10):1243-50. Epub > 2006 Sep 19. > A time- and cost-efficient system for high-level protein production in > mammalian cells. > Aricescu AR, Lu W, Jones EY. > PMID: 17001101 > > Nat Protoc. 2018 Dec;13(12):2991-3017. doi: 10.1038/s41596-018-0075-9. > Lentiviral transduction of mammalian cells for fast, scalable and > high-level production of soluble and membrane proteins. > Elegheert J, Behiels E, Bishop B, Scott S, Woolley RE, Griffiths SC, > Byrne EFX, Chang VT, Stuart DI, Jones EY, Siebold C, Aricescu AR. > PMID: 30455477 > > Hope that helps, > Tomas > > > Dr. Tomas Malinauskas > University of Oxford > Wellcome Centre for Human Genetics > Division of Structural Biology > Roosevelt Drive > Oxford OX3 7BN > United Kingdom > to...@strubi.ox.ac.uk > tomas.malinaus...@gmail.com > >> On Fri, Jan 24, 2020 at 11:50 PM Gloria Borgstahl <gborgst...@gmail.com> >> wrote: >> >> Hello CCP4-ers, >> I was wondering what people have found to be the best human cell line >> expression system for making a large quantity of purified recombinant >> protein. >> Any information and protocols would be greatly appreciated. >> Happy 2020, Gloria >> >> ________________________________ >> >> To unsubscribe from the CCP4BB list, click the following link: >> https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 > > ######################################################################## > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
