I want to clone a secretory protein from H. pylori and transformation will be done in E. coli BL21. The protein has a 33 aa signal sequence in its N terminal and auto transporter domain at C terminus. I'm just wondering whether these domains are necessary during expression in E.coli BL21. or does adding His tag at N or C terminals will hinder with signal or autotransporter domains expression. Kindly help me.
Generally, there is no problem with His-tags on exported proteins in bacterial expression system. If you add the tag N-terminally, you would of course have to add it after the export signal sequence and start it with an amino acid that is compatible with proper signal peptidase processing. Whether an N-terminal or C-terminal His tag could interfere with the function of your protein depends very much on the specific protein - it only rarely does, but it cannot be excluded. That is something you just have to try out.
Generally, there is no problem with His-tags on exported proteins in bacterial expression system. If you add the tag N-terminally, you would of course have to add it after the export signal sequence and start it with an amino acid that is compatible with proper signal peptidase processing. Whether an N-terminal or C-terminal His tag could interfere with the function of your protein depends very much on the specific protein - it only rarely does, but it cannot be excluded. That is something you just have to try out.
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