pET24dTEV – DDX3pfSTREPHIS
# 44
Name: Bernardo Rodamilans
Gene/Protein
DDX3 fragment (1347bp)
Organism
Human
Plasmid
pETM-11 similar to pET24d (KanR)
Restriction sites
BspHI, BamHI, NcoI
Concentration
452 ng/uL
Bacteria - Expression
BL21
Bacteria - Molecular biology
XL1Blue
Date: 24.03.04
Comments
The first amino acids are DDIPV. The
last aa are GGFGAR. The protein has
a 6HIStag at the n-terminal that can be
cut with TEV protease and also a
STREPtag at its c-terminal with a SA
link.
The insert has been cut with BspHI
and BamHI.
The vector has been cut with NcoI and
BamHI.
BspHI and NcoI produce compatible
cohesive ends generating recleavable
ligation products. However, once insert
and vector are ligated, both sites
disappear.
To check if ligation worked well, NcoI
and BamHI can be used. Since there is
an NcoI site in the DDX3 gene, two
fragments will be generated, one with
7000bp, and the other one 500bp long.
Map
BamHI
NcoI
PET24d-DDX3pfSTREPHIS
7400bp