pET28a – DDX3 c-terminal HIStag
# 14
Name: Bernardo Rodamiláns
Date: 18.03.03
Gene/Protein
DDX3 (2007bp)
Comments
The protein has a 6HIStag at the cterminal.
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
6685bp, and the other one 595bp long.
Organism
Human
Plasmid
PET28a (KanR)
Restriction sites
BspHI, BamHI, NcoI
Concentration
257.7 ng/uL
Bacteria - Expression
BL21
Bacteria - Molecular biology
XL1Blue
Map
BamHI 7079
NcoI 6484
PET28aDDX3cterHIS
7280bp