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Table 1 Effect of MtTOP1 substitutions on the complementation of topAtsfunction in E. coli AS17 at the non-permissive temperature

From: The DNA relaxation activity and covalent complex accumulation of Mycobacterium tuberculosis topoisomerase I can be assayed in Escherichia coli: application for identification of potential FRET-dye labeling sites

Plasmid

Complementation ratio

pLIC vector

3.9 × 10-5 ± 3.3 × 10-5

pLIC-ETOP

0.58 ± 0.21

pLIC-MTOP

0.53 ± 0.24

pLIC-MTOP-Y342A

1.1 × 10-4 ± 1.1 × 10-4

pLIC-MTOP-G116S

3.3 × 10-5 ± 2.0 × 10-5

pLIC-MTOP-M136C

2.7 × 10-4 ± 4.1 × 10-4

pLIC-MTOP-H139C

0.19 ± 0.15

pLIC-MTOP-T142C

0.097 ± 0.029

pLIC-MTOP-L170C

0.32 ± 0.16

pLIC-MTOP-Y174C

0.18 ± 0.06

pLIC-MTOP-K524C

0.33 ± 0.17

  1. Serial dilutions of overnight cultures of E. coli AS17 transformants were plated on LB plates with kanamycin and incubated for 2 days at either 30°C or 42°C. Colony counts from 42°C incubation were divided by colony counts from 30°C to obtain the complementation ratios. The results represent the average and standard deviation from at least three measurements.
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BMC Biochemistry

ISSN: 1471-2091

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