EXPRESSION OF THE CBS MUTANT ALLELES IN E. COLI

BACK

 
 

cDNA NT change

Protein change

GENE NT change

GENE exin

Residual activity

% of mean WT

172 C>T R58W 172 C>T EX1
1.3
233 C>G P78R 3600 C>G EX2
12.6-60.9
253G>A G85R 3620G>A EX2
0
302 T>C L101P 3669T>C EX2
0
306 G>C K102N 3673 G>C EX2
33.3-38.7
325 T>C C109R 5824 T>C EX3
0
341 C>T A114V 5840 C>T EX3
0-54.5
374 G>A R125Q 5873 G>A EX3
1.9
374 G>C R125P 5873 G>C EX3
n.d.
429 C>G I143M 5928 C>G EX3
0-4
434 C>T P145L 5933 C>T EX3
0.00
442 G>A G148R 5941G>A EX3
0
442insG N149fsX187 5941insG EX3
0
456 C>G I152M 6503 C>G EX4
0.3
494 G>A C165Y 6541 G>A EX4
1-2
526 G>A E176K 6573 G>A EX4
0
539 T>C V180A 6681 T>C EX5
3
572 C>T T191M 6714 C>T EX5
1
684 C>A N228K 6940 C>A EX6
0
684 C>G N228K 6940 C>G EX6
0
694 C>G H232D 6950 C>G EX6
7.11
727 C>T Q243X 6983 C>T EX6
0
833 T>C I278T 9122 T>C EX8
0-1.5
904 G>A E302K 9193 G>A EX8
3.9-5.5
919 G>A G307S 9208 G>A EX8
0
919 G>A G307S 9208 G>A EX8
0.00
1006 C>T R336C 9851 C>T EX9
0.00
1039 G>A G347S 9884 G>A EX9
0
IVS9+1 G>T del ex 9, T318fsX330 9885 G>T IN9
0-3
1111 G>A V371M 11717 G>A EX10
1.1
IVS11-2 A>C del ex 12 13217 A>C IN11
0.00
1265 C>T P422L 13260 C>T EX12
100
1304T>C I435T 13299T>C EX12
100
1316 G>A R439Q 13311 G>A EX12
30
1321 A>T K441X 13316 A>T EX12
128.9
1330 G>A D444N 13325 G>A EX12
75.9
1397C>T S466L 13972C>T EX13
100

The mutations were introduced into an expression plasmid pHCS3 derived from pKK388.1. CBS activity was measured in E.coli lysates as described (Kozich and Kraus, 1992; de Franchis et al., 1994 ). The activity of E144K, R439Q, T353M mutants were determined after expression in expression system derived from plasmid pT7.7 (Dawson et al., 1997)



Last Updated on 08/04/2009 by Krauslab.