PKA alpha expression with Chaperones
(Redirected from PKA alpha expression)
Medium: LB/Kan/Chl
Screening of PKA alpha expression with chaperones
| # | Plasmid | Chaperone | Promoter | Inducer | Resistant Marker |
| 1 | pG-KJE8 | dnaK-dnaJ-grpE | araB | L-Arabinose | Cm |
| groES-groEL | Pzt1 | Tetracyclin | |||
| 2 | pGro7 | groES-groEL | araB | L-Arabinose | Cm |
| 3 | pKJE7 | dnaK-dnaJ-grpE | araB | L-Arabinose | Cm |
| 4 | pG-Tf2 | groES-groEL-tig | Pzt1 | Tetracyclin | Cm |
| 5 | pTf16 | tig | araB | L-Arabinose | Cm |
Should induce with 0.02% to 0.2% L-Arabinose (and + Tetracyclin )
weight of wet cell pellet from 1 L culture
| pellet wt | PKA yeild | |
| pG-KJE8 |
4.3 | |
| pGro7 |
3.9 | |
| pKJE7 |
3.6 | |
| pG-Tf2 |
1.4 | |
| pTf16 |
4.6 |
| 2012/11/28 |
pellet wt from 2 L | PKA yeild |
| pG-KJE8 |
4.0 | |
| pGro7 |
5.1 | |
| pKJE7 |
4.5 |
Expression screening
1 pTf16
2 pG-KJE8
3 pGro7
4 pG-Tf2
5 pKJE7
6 pTf16
7 pG-KJE8
8 pGro7
9 pG-Tf2
10 pKJE7
Looks like pG-KJE8 (tet+L-arabinose induction) has better expression
--Ruili.su 17:45, 18 March 2011 (EST)
PKA purification using gel-filtration column