pET28a – DDX3pf
# 41
Name: Bernardo Rodamiláns
Gene/Protein
DDX3 fragment (1311bp)
Organism
Human
Plasmid
PET28a (KanR)
Restriction sites
BspHI, BamHI, NcoI
Concentration
470 ng/uL
Bacteria - Expression
BL21
Bacteria - Molecular biology
XL1Blue
Date: 16.02.04
Comments
This is a fragment of the DDX3protein
according to a proteolytic assay. The
first amino acids are DDIPV. The last
aa are GGFGAR. The protein has a
6HIStag at the c-terminal.
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
6000bp, and the other one 400bp long.
Map
BamHI
NcoI
PET28aDDX3 pf
6400bp