2115 2203 with chaperones
Use chaperones to improve Flp expression
[edit]5/27/2014
Test chaperone on -yFLP expression
| Lane |
Name |
Protein Name |
Clone ID |
Host |
Induction |
Purification |
sample |
| 1 |
pT7A3-iFLP 9.76 / 52.8 |
iFLP |
2115 |
15 ml, Tuner(DE3)-pG-KJE8 |
0.2% arabinose+5ng/ml Tetracycline 0.3 mM IPTG 30oC o/n |
Ni beads |
S |
| 2 |
1 5 ml, Tuner(DE3)-pGro7 |
0.2% arabinose+ 0.3 mM IPTG 30oC o/n |
Ni beads |
S | |||
| 3 |
15 ml, Tuner(DE3)-pKJE7 |
0.2% arabinose+ 0.3 mM IPTG 30oC o/n |
Ni beads |
S | |||
| 4 |
15 ml, Tuner(DE3)-pGTf2 |
5ng/ml Tetracycline 0.3 mM IPTG 30oC o/n |
Ni beads |
S | |||
| 5 |
15 ml,
Tuner(DE3)-pTf16 |
0.2% arabinose+ 0.3 mM IPTG 30oC o/n |
Ni beads |
S | |||
| 6 |
Fermentas PageRuler Plus |
|
| ||||
| 7 |
pT7A3-iFLP
9.76 / 52.8 |
iFLP |
2115 |
15 ml,
Tuner(DE3)-pG-KJE8
|
0.2% arabinose+5ng/ml Tetracycline
0.3 mM IPTG 30oC o/n |
Ni beads |
p |
| 8 |
1 5 ml,
Tuner(DE3)-pGro7
|
0.2% arabinose+
0.3 mM IPTG 30oC o/n |
Ni beads |
p | |||
| 9 |
15 ml,
Tuner(DE3)-pKJE7
|
0.2% arabinose+
0.3 mM IPTG 30oC o/n |
Ni beads |
p | |||
| 10 |
15 ml,
Tuner(DE3)-pGTf2
|
5ng/ml Tetracycline
0.3 mM IPTG 30oC o/n |
Ni beads |
p | |||
| 11 |
15 ml,
Tuner(DE3)-pTf16 |
0.2% arabinose+
0.3 mM IPTG 30oC o/n |
Ni beads |
p | |||
| 12 |
| Lane |
Name |
Protein Name |
Clone ID |
Host |
Induction |
Purification |
sample |
| 1 |
pCold-MBP-Tat-NLS-iFLP-6H 9.03 / 96.9 |
MBP-Tat-NLS-iFLP-6H |
2203 |
15 ml, Tuner(DE3)-pG-KJE8 |
0.2% arabinose+5ng/ml Tetracycline 0.3 mM IPTG 30oC o/n |
Ni beads |
S |
| 2 |
1 5 ml, Tuner(DE3)-pGro7 |
0.2% arabinose+ 0.3 mM IPTG 30oC o/n |
Ni beads |
S | |||
| 3 |
15 ml, Tuner(DE3)-pKJE7 |
0.2% arabinose+ 0.3 mM IPTG 30oC o/n |
Ni beads |
S | |||
| 4 |
Fermentas PageRuler Plus |
|
| ||||
| 5 |
9.03 / 96.9 |
MBP-Tat-NLS-iFLP-6H |
2203 |
15 ml,
Tuner(DE3)-pGTf2
|
5ng/ml Tetracycline
0.3 mM IPTG 30oC o/n |
Ni beads |
S |
| 6 |
15 ml,
Tuner(DE3)-pTf16 |
0.2% arabinose+
0.3 mM IPTG 30oC o/n |
Ni beads
|
s | |||
| 7 |
15 ml,
Tuner(DE3)-pG-KJE8
|
0.2% arabinose+5ng/ml Tetracycline
0.3 mM IPTG 30oC o/n |
Ni beads |
p | |||
| 8 |
1 5 ml,
Tuner(DE3)-pGro7
|
0.2% arabinose+
0.3 mM IPTG 30oC o/n |
Ni beads |
p | |||
| 9 |
15 ml,
Tuner(DE3)-pKJE7
|
0.2% arabinose+
0.3 mM IPTG 30oC o/n |
Ni beads |
p | |||
| 10 |
15 ml,
Tuner(DE3)-pGTf2
|
5ng/ml Tetracycline
0.3 mM IPTG 30oC o/n |
Ni beads |
p | |||
| 11 |
15 ml,
Tuner(DE3)-pTf16 |
0.2% arabinose+
0.3 mM IPTG 30oC o/n |
Ni beads |
p | |||
| 12 |
Conclusion: Chaperones do not enhance protein expression. Protein is also insoluble