Improve Research Reproducibility A Bio-protocol resource
- Home
- Protocols
-
Concise Method
Cloning of various constructs used in this study
Request a Protocol
Ask a
question
Favorite
EhGEF was cloned in amoebic expression vector pEhHYG-tetR-O- CAT by replacing CAT gene with the gene of interest in the cassette using double digestion by KpnI and BamHI either in sense or antisense orientations. EhGEF was cloned into pEh-Neo-GFP vector under Xho1 and BamH1 sites so that GFP comes at N-terminus of protein. HA-tagged GEF was cloned in pEh-Neo-3HA vector under SmaI and XhoI sites. The ΔEhGEF was generated by introducing stop codon at 121 position (Alanine to stop codon) by site directed mutagenesis.
Copyright and License information: ©2021 Bharadwaj et al ©2021 Carruthers, Petri, Jr.This is an open access article distributed under the terms of the , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. ©2021 Carruthers, Petri, Jr.This is an open access article distributed under the terms of the , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Do you have any questions about this protocol?
Post your question to gather feedback from the community. We will also invite the authors of this article to respond.
Post a Question0 Q&A
