Tools for genetic manipulation of the plant growth-promoting bacterium Azospirillum amazonense

BMC Microbiology

Table 2 Primers utilized in this work

Primers	Sequence	Annealing temperature	Amplicon length (bp)	Purpose
glnB_sfint	CGCCGCGATACAGCTCGGTATG	57°C	2108	glnB region isolation
revsf_glnBint	GATGGACGATCAGTTGGTCGA	57°C	2108	glnB region isolation
glnA_aa_do	ACGGTCGGCACTTCCTTCAG	53°C	1621	glnB region isolation
pglnB_up_BamHI	CGGGATCCTCGTCGAAGCTGAAGGTCAT	55°C	727	glnB promoter amplification
pglnB_do_NcoI	GATCTTTTCCATGGCTTACGGC	55°C	727	glnB promoter amplification
pglnK_up_BamHI	CGGGATCCTTGTCCAGTGCCACGCTCAT	55°C	328	glnK promoter amplification
pglnK_do_NcoI	CACGAGCTCCATGGGTAGTCC	55°C	328	glnK promoter amplification
Kglndel_A _EcoRI	ATGAATTCCAATGCACAGGGTGCGTA	55°C	574 (AB) and 1111 (AD)	glnK mutagenesis
Kglndel_B	CCCATCCACTAAACTTAAACACGCCACCACGAGCTTCAT	55°C	574	glnK mutagenesis
Kglndel_C	TGTTTAAGTTTAGTGGATGGGATGACCATCGCCGACGCG	55°C	558	glnK mutagenesis
Kglndel_D_BamHI	CGGGATCCCGATGGTGGGCGGATATTTG	55°C	558 (CD) and 1111 (AD)	glnK mutagenesis
glnK_NdeI_up	GGACTACATATGAAGCTCGTGGTG	60°C	361 (wt) or 121 (mut)	glnK mutagenesis verification
glnK_BamHI_do	CGTCACGGGATCCTCATAAGGC	60°C	361 (wt) or 121 (mut)	glnK mutagenesis verification
conf_glnK_up	GCCCCCTCCAGGATCTTC	55°C	1522 (wt) or 1282 (mut)	glnK mutagenesis verification
conf_glnK_do	GGGTAAAATGCCCTTGTCCA	55°C	1522 (wt) or 1282 (mut)	glnK mutagenesis verification

Underline - restriction sites; Bold - sequence tag; wt - wild-type; mut - mutant; AB - amplification using the primers KglndelA_EcoRI and KglndelB; AD - amplification using the primers Kglndel_A_EcoRI and Kglndel_D_BamHI; CD - amplification using the primers KglndelC and Kglndel_D_BamHI

ISSN: 1471-2180