PKA alpha expression with Chaperones

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