Do we express our recombinant protein into TOP10 instead of BL21(DE)? Is this affects its protein expression?
Hi there,
If you were using BL21(DE3) at first it means your gene of interest is under the control of T7 promoter which requires T7 RNA polymerase for efficient transcription. If using TOP10 cells (which are actually optimized for cloning purpose and plasmid amplification) instead with the same plasmid you won't be able to produce any protein as TOP10 doesn't code for T7 RNA polymerase.
If you aim to express your protein of interest, then BL21DE3 (Competent E. coli with expression vector system) would be more suitable than competent TOP10 because TOP10 cells are mostly used for plasmid constructing.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3732866/
Hi there,
If you were using BL21(DE3) at first it means your gene of interest is under the control of T7 promoter which requires T7 RNA polymerase for efficient transcription. If using TOP10 cells (which are actually optimized for cloning purpose and plasmid amplification) instead with the same plasmid you won't be able to produce any protein as TOP10 doesn't code for T7 RNA polymerase.
- Badges
- Science method
- Similar topics
- Biological Science
- Genetics
- Gene Expression
- Protein Expression