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Table 3 Bacterial strains and plasmids used in this study.

From: High-level chromate resistance in Arthrobactersp. strain FB24 requires previously uncharacterized accessory genes

Strain or plasmid

Description

Reference

Arthrobacter

  

   FB24

CrR

[6]

   D11

CrS derivative of FB24

This work

E. coli

  

   JM110

dam - dcm -

Stratagene

Plasmids

  

pAOWA10128

7.3 kb insert in pMCL200 obtained from DOE-JGI. Contains Arth_4248-Arth_4254.

DOE-JGI

pBluescript II SK+

3.0 kb, ApR, lacZ, used for sublconing inserts prior to ligation into pART2.

Promega

pART2

4.6 kb, KmR, pCG100 ori, ColE1 ori, vector for expression in Arthrobacter

[55]

pKH11

10.6 kb PCR product from FB24 plasmid 3 (CP000457) containing Arth_4247-4255 in pBluescript II SK+

This worka

pKH12

Insert from pKH11 cloned into pART2

This work

pKH21

7.3 kb insert from pAOWA10128 in pBluescript II SK+

This work

pKH22

Insert from pKH21 cloned into pART2

This work

pKH32

3.7 kb EcoRI-KpnI fragment from pKH21 cloned into pART2. Contains Arth_4248-4249.

This work

pKH42

3.8 kb XhoI-BglII fragment from pKH21 cloned into pART2. Contains Arth_4251-Arth_4254.

This work

pKH52

8.3 kb insert from MluI-BglII digest of pKH11 to delete Arth_4252 and Arth_4252 cloned into pART2

This work

pKH62

pKH22 digested with SfiI to delete Arth_4249-Arth_4252.

This work

pKH72

pKH12 digested with ScaI and XbaI to delete Arth_4247.

This work

  1. aA schematic of each construct is presented in Figure 3.