. Author manuscript; available in PMC: 2012 Oct 4.

Published in final edited form as: J Biol Chem. 2005 Oct 31;281(2):1119–1127. doi: 10.1074/jbc.M509421200

TABLE 2. Screening results for constitutively active CRP variants randomly altered at Th¹²⁷ and Ser¹²⁸.

The codons at positions 127 and 128 of CRP were simultaneously randomized, and the the pool of randomized clones (crp gene in pEXT20) was transformed into strain UQ3811 and screened on LC agar plates containing 50 μg/ml ampicillin, 20 μg/ml 5-bromo-4-chloro-3-indolyl-β -D-galactopyranoside (X-gal), and 0 or 100 μM IPTG.

Screened without IPTG					Screened with 100 μM IPTG
CRP	Position 127		Position 128		CRP	Position 127		Position 128		CRP	Position 127		Position 128
CRP	Codon	Amino acid	Codon	Amino acid	CRP	Codon	Amino acid	Codon	Amino acid	CRP	Codon	Amino acid	Codon	Amino acid
LB-1^a	TTA	Leu	ATA	Ile	DB-1'^b	ATA	Ile	ATT	Ile	LB-1'^c	ATG	Met	ACA	Thr
LB-2	CTG	Leu	ATT	Ile	DB-2'	ATG	Met	GTA	Val	LB-2'	TTG	Leu	CAA	Gln
LB-3	CTG	Leu	ATA	Ile	DB-3'	CTT	Leu	GTG	Val	LB-3'	TTG	Leu	CTA	Leu
LB-4	CTG	Leu	ATT	Ile	DB-4'	TTG	Leu	GTC	Val	LB-4'	ACA	Thr	ATA	Ile
LB-5	TTG	Leu	ATC	Ile	DB-5'	CTA	Leu	GTG	Val	LB-5'	CTG	Leu	ACT	Thr
LB-6	CTG	Leu	ATA	Ile	DB-6'	CTT	Leu	GTG	Val	LB-6'	CTC	Leu	ATG	Met
LB-7	TTA	Leu	ATT	Ile	DB-7'	ATG	Met	GTA	Val	LB-7'	CTA	Leu	ACT	Thr
LB-8	TTA	Leu	ATA	Ile	DB-8'	CTA	Leu	AAG	Lys	LB-8'	ATG	Met	ATG	Met

Light blue (2.3% of the total population).

Dark blue (3.4% of the total population).

Light blue (8.5% of the total population).

TABLE 2. Screening results for constitutively active CRP variants randomly altered at Th127 and Ser128.

TABLE 2. Screening results for constitutively active CRP variants randomly altered at Th¹²⁷ and Ser¹²⁸.