Hi Song, I think Lam4 STARTkin domain could be a good choice for you. It expresses quite well in E.coli and migrate nicely in the SDS-PAGE gel. The protein is quite stable and molecular weight of this protein is 20.5KDa. You can find the structure and sequence that have used here http://www.rcsb.org/structure/5YQJ. Best wishes, Mohammad Kawsar Manik Kobe lab, The University of Queensland
From: CCP4 bulletin board <[email protected]> on behalf of "Tan, Song" <[email protected]> Reply-To: "Tan, Song" <[email protected]> Date: Monday, 7 January 2019 at 9:25 am To: "[email protected]" <[email protected]> Subject: [ccp4bb] looking for 10-20 kDa proteins that express well and migrate appropriately on SDS-PAGE 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 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
