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Table 5 Strains, plasmids and primers used in this experiment

From: The Mesorhizobium huakuii transcriptional regulator AbiEi plays a critical role in nodulation and is important for bacterial stress response

Strains

Description

Reference, Source, Sequence

M. huakii 7653R

Wild type, Nod+ on Astragalus sinicus

[45]

HKabiEi

7653R abiEi:pk19mob, Strr Neor

This study

HKabiEi(pBBRabiEi)

Strain HKabiEi harboring plasmid pBBRabiEi

This study

HKabiEi(pBBR1MCS-5)

Strain HKabiEi harboring plasmid pBBR1MCS-5

This study

Plasmids

 pETAbiE

AbiE_F/AbiE_R PCR product in pET-28a(+), Kmr

This study

 pETAbiEii

AbiE_F/AbiEii_R PCR product in pET-28a(+), Kmr

This study

 pK19mob

pK19mob pUC19 derivative lacZ mob Kmr

[46]

 pRK2013

Helper plasmid for mobilizing plasmids Kmr

[47]

 pKabiEi

abiEiUP/abiEiLW PCR product in pK19mob, Kmr

This study

 pBBR1MCS-5

lacPOZ′ mob, broad host range, Gmr

[48]

 pBBRabiEi

cabiEi_F/cabiEi_R PCR product in pBBR1MCS-5, Gmr

This study

Primera

 AbiE_F

Sense primer used for AbiE system and abiEii gene expression

TTTGGATCCATGTCCTTGGTTGAACCCGA

 AbiE_R

Antisense primer used for AbiE system expression

TTTAAGCTTGTGCAGGTCGTAGTAGTGGC

 AbiEii_F

Antisense primer used for MCHK_RS33185

(abiEii) expression

TTTGGATCCGTGAGCACCGACGCCTATCG

 abiEiUP

Sense primer for MCHK_RS33180

(abiEi) mutation

TTTAAGCTTGAACCCGACAGCGATCTCCG

 abiEiLW

Antisense prime for MCHK_RS33180 (abiEi) mutation

TTTTCTAGAACTTGGCCTCGCTCGATGAG

 abiEipimap

Mapping PCR primer for abiEi

TGACTCCAGCGCAGCCGTCA

 cabiEi_F

Sense primer for abiEi complementation

TTTAAGCTTGCCACCGACCTTTTATCCTG

 cabiEi_R

Antisense primer for abiEi complementation

TTTTCTAGAGCCCTTGAACAGCAGCCGCG

 pK19A

pK19mob mapping primer

ATCAGATCTTGATCCCCTGC

 pK19B

pK19mob mapping primer

GCACGAGGGAGCTTCCAGGG

 Q16SrRNAF

Sense primer for qRT-PCR of 16S rDNA

AACTGAGATGGCTTTTGGAG

 Q16SrRNAR

Antisense primer for qRT-PCR of 16S rDNA

GGATGACGTCAAGTCCTCAT

 Q32450F

Sense primer for qRT-PCR of MCHK_RS32450

TGGCGAGGTTACCGTACTCA

 Q32450R

Antisense primer for qRT-PCR of MCHK_RS32450

AGCGTGTCTGAGCCGGCAAA

 Q32465F

Sense primer for qRT-PCR of MCHK_RS32465

GCTCTACCAGAACCGCTATC

 Q32465R

Antisense primer for qRT-PCR of MCHK_RS32465

TCGTACAACTCGTAGCATTT

 Q32345F

Sense primer for qRT-PCR of MCHK_RS32345

CAACATCCATCAGATCACGA

 Q32345R

Antisense primer for qRT-PCR of MCHK_RS32345

TGGCGCATTTCTCCGTCTTT

 Q31505F

Sense primer for qRT-PCR of MCHK_RS31505

CAATGGAGTTTCTCAATGGA

 Q31505R

Antisense primer for qRT-PCR of MCHK_RS31505

TATAGAGAAGCAGAGGCGTT

 Q31555F

Sense primer for qRT-PCR of MCHK_RS31555

TGATCAGGCAGAACCACGGC

 Q31555R

Antisense primer for qRT-PCR of MCHK_RS31555

ATCAAGGTCCGTACCCGTTA

 Q31330F

Sense primer for qRT-PCR of MCHK_RS31330

TCGGCTCAGAATATCCTCGT

 Q31330R

Antisense primer for qRT-PCR of MCHK_RS31330

CGAAATGAAGTCGCTTAGTA

 Q31340F

Sense primer for qRT-PCR of MCHK_RS31340

ATTGCCAGCGCCAGCCAGGG

 Q31340R

Antisense primer for qRT-PCR of MCHK_RS31340

GGCCGGGCTGCTCTATCTCG

QabiEiF

Sense primer for qRT-PCR of abiEi

TCAAGAGCTTCGCCGGATCG

QabiEiR

Antisense primer for qRT-PCR of abiEi

ACGGATCACGGCGCGATAGT

QabiEiiF

Sense primer for qRT-PCR of abiEii

GGCCCGCTCAATGAGTTCCT

QabiEiiR

Antisense primer for qRT-PCR of abiEi

TCTGATACCAAAGCAGCAGG

  1. aRestriction sites in primer sequences are underlined