Off the top of my head, for your mass range with gel behavior that you ask for:
Human or mouse Synuclein (not folded, very heat stable, incredibly soluble) - interesting reaction with SDS but runs fine on the gel T4 Lysozyme with a double mutation (expresses mega-loads per gram of cell paste) Ferredoxin Rubredoxin cytochrome b562 Ubiquitin the list goes on and on... As to polycistronic stuff - it may be easier to mix lysates than to mess around with multiple expression cassettes - in case you'd want to change the mix later... just my two cents. Artem - Cosmic Cats approve of this message On Sun, Jan 6, 2019 at 6:15 PM Tan, Song <[email protected]> wrote: > Dear Colleagues, > > As a followup to my lab’s plasmids for making DNA molecular weight markers > <http://www.personal.psu.edu/sxt30/projects_DNA_ladders.html> for about > one cent per lane (Henrici et al, Scientific Reports, 7:2438, 2017 > <https://www.nature.com/articles/s41598-017-02693-1>; plasmids are > available from Addgene), we are working on a similar project to produce > protein MW markers in E. coli. We are having problems finding 10-20 kDa > proteins that express solubly at high levels in E. coli and that migrate > appropriately on SDS-PAGE. We have eliminated multiple proteins including > thioredoxin and DHFR that express very well but that migrate anomalously > slowly on SDS-PAGE. > > So we are looking for proteins which fulfill the following criteria: > > 1. MW = 8-10 kDa or 18-20 kDa, > 2. Express as soluble protein in E. coli, > 3. Can be purified from E. coli with yields of greater than 10 mg/L > culture, > 3. Migrate appropriately on SDS-PAGE. > > We hope to create a simple and inexpensive system for producing protein MW > ladders, and to distribute the plasmids through Addgene. Plan is to > coexpress multiple proteins using our polycistronic expression system > <http://www.personal.psu.edu/sxt30/projects_polycistronic.html> to make > the production process simpler. > > If you know of proteins that fulfill all of the above criteria, please do > drop me a line. > > Regards and thanks, > > Song > > --------------------------------------- > > Dr. Song Tan > Professor of Biochemistry & Molecular Biology > Center for Eukaryotic Gene Regulation > Dept of Biochemistry & Molecular Biology > 108 Althouse Laboratory (office in 468A North Frear Lab) > Penn State University > University Park, PA 16802 > email: [email protected] <[email protected]> > phone: 814-865-3355 fax: 814-863-7024 > web: http://www.personal.psu.edu/sxt30 > > > ------------------------------ > > 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
