Hi Dhiraj, For transient expression in HEK293 cells in suspension, I generally use pcDNA3.1 which also uses a CMV promoter. Yields range from nothing to 50mg/l, depending entirely on what it is you're making.
I would suggest that maybe the promoter is not at fault, and perhaps the product is. Tweaking construct boundaries and optimizing transfection conditions may help. Good luck, Dave -- Dr David C. Briggs Senior Laboratory Research Scientist Signalling and Structural Biology Lab The Francis Crick Institute London, UK == about.me/david_briggs ________________________________ From: CCP4 bulletin board <[email protected]> on behalf of Srivastava, Dhiraj <[email protected]> Sent: Thursday, June 24, 2021 9:42:32 PM To: [email protected] <[email protected]> Subject: [ccp4bb] Eukaryotic protein expression External Sender: Use caution. Hi all I am trying to express my protein in HEK293 cells for crystallization purpose but getting poor expression. I am using cmv promoter. Which promoter/vector people use to get good expression in eukaryotic expression system? Thank you Dhiraj ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1<https://eur03.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.jiscmail.ac.uk%2Fcgi-bin%2FWA-JISC.exe%3FSUBED1%3DCCP4BB%26A%3D1&data=04%7C01%7C%7C63244fd02e074ee429f408d937509d24%7C4eed7807ebad415aa7a99170947f4eae%7C0%7C1%7C637601641663192554%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&sdata=eD0GBPXnEwomdM9DOF7Gki85xqBnFGLzsYkWiuY2mmg%3D&reserved=0> The Francis Crick Institute Limited is a registered charity in England and Wales no. 1140062 and a company registered in England and Wales no. 06885462, with its registered office at 1 Midland Road London NW1 1AT ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
