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Construction of A. brasilense cheA2::tet cheA3::Tn5 strain.

LO Lindsey O’Neal
JG Jessica M. Gullett
AA Anastasia Aksenova
AH Adam Hubler
AB Ariane Briegel
DO Davi Ortega
AK Andreas Kjær
GJ Grant Jensen
GA Gladys Alexandre
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To generate the cheA2::Tet cheA3::Tn5 strain, a cheA2 knockout insertion was produced using a pKNOCK suicide vector carrying an internal fragment of cheA2 and introduced into the cheA3::Tn5 strain by mating. pKNOCKCheA2 was generated by amplifying an internal fragment of cheA2 (NCBI accession no. ALJ38472.1) using the primers listed in Table S2. The pKNOCK vector and the PCR fragments were digested with SmaI, ligated, and transformed into competent E. coli TOP10 cells. E. coli TOP10(pKNOCKCheA2) then was used as a donor in conjugation with the A. brasilense cheA3::Tn5 derivative (20). Disruptants were selected on MMAB with tetracycline (10 mg/ml) and confirmed using colony PCR.

Copyright and License information:  ©2019 O’Neal et al.
This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

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