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]<mailto:[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
