Table 2.

List of primers used in this study^a

No.	PCR amplicon	Size (bp)	Oligonucleotide sequence
1	Genomic library clone carrying kshA1	227	KshA1Rho-F, GCCGCTGCAAGAACATCCCGTAC
			KshA1Rho-R, GATGATCTCGCGGCAGTTGGTGTC
2	Genomic library clone carrying kshA3	547	KshA3Rho-F, CGACCTGTCACAGGGCGAGAT
			KshA3Rho-R, TACGGATTCGGTCTGGTACC
3	Genomic library clone carrying kshA3	161	KshA5Rho-F, CGCCTGGACGACCCTCGAACGCA
			KshA5Rho-R, AGTGGGAGCCCTTGATGCGCAG
4	Internal fragment ro04690 homolog (screening genomic library)	361	ro04690DEG-F, CACACCGG(C/T)GAG(A/G)T(C/G)GCG(A/T)C(C/G)ATG
			ro04690DEG-R, TCTGCAGCGGCAT(C/G)(C/G)(C/G)(A/C)AGCGG
5	Internal fragment fadE26 homolog (screening genomic library)	670	fadE26DEG-F, (C/G)(A/G)TCAACGG(A/C)CAGAAGATGTGG
			fadE26DEG-R, A(C/T)(C/T)TCGTTGGT(G/T)CCGCCGCCGAA
6	fadD19DSM43269 gene (heterologous expression in pET15b)	1,679	fadD19exp-F, TCATATGGCCCTAAACATCGCAGACC
			fadD19exp-R, TAGATCTCTATCCCGTCGCCGCGGCGCCG
7	fadD19DSM43269 gene (confirmation ΔfadD19 mutant)	527 (wt, 1,278)	fadD19Contr-F, GTGAAGACCGTCGTGGTGGTAG
			fadD19Contr-R, GTTGCCGTCGCCGTTGTTGTAG
8	fadD1943269 gene (complementation ΔfadD19 mutant)	1,673	COMPfadD19-F, GGCCCTAAACATCGCAGACCTC
			COMPfadD19-R, CGGTACCCTATCCCGTCGCCGCGGCGCCG
9	Internal fragment fadD17DSM3269 (disruption construct)	826	fadD17-F, GACATCGCGCTCGCCGATTGCC
			fadD17-R, TGTAGTAGCCGTCGAACAGACC
10	fadD17DSM43269 (confirmation fadD17 disruption)	1,027	pK18Contr-F, AATGCAGCTGGCACGACAGGTT
			fadD17Contr-R, CGAGCATCCTGCGCGAACTCGG
11	Internal fragment fadE27DSM3269 (disruption construct)	753	fadE27-F, GCAGCATCGTCGCGGACATC
			fadE27-R, GGAGCCGAGCAGGAACTCGT
12	fadE27DSM43269 (confirmation fadE27 disruption)	1,510	fadE27Contr-F, GCAGCATCGTCGCGGACATC
			pK18Contr-R, CTGCGTGCAATCCATCTTGTTC
13	Upstream region ro04690DSM43269 (deletion construct)	1,247	ro04690UP-F, AGCGCCGACGACATCTACATCC
			ro04690UP-R, TCATATGCGTGAATCCGAAGATCGGATAC
14	Downstream region ro04690DSM43269 (deletion construct)	1,236	ro04690DOWN-F, TCATATGGAGATCATGGCCGAACTCGTC
			ro04690DOWN-R, CTGCAGGATCACGGCAACGAC
15	ro04690DSM43269 gene (confirmation Δro04690 mutant)	129 (wt, 1,113)	ro04690Contr-F, ATGCAGACCGGCCTCAGCAAGA
			ro04690Contr-R, CTAGCGGGCCTCGTTCAGCCGTTC
16	Upstream region fadE26DSM43269 (deletion construct)	1,234	fadE26UP-F, TCGAGCAGTTCGGTAACGGTGAG
			fadE26UP-R, AGATCTCGAGGTCCAGCGCGACATCATC
17	Downstream region fadE26DSM43269 (deletion construct)	1,230	fadE26DOWN-F, TTAGATCTTTCCTCGCGGAGGGCTTGTTGC
			fadE26DOWN-R, GGCGAGATAGGCGACCAGATTC
18	fadE26DSM43269 gene (confirmation ΔfadE26 mutant)	520 (wt, 1,593)	fadE26Contr-F, GCTTCACGCACGGCGTCCTCAC
			fadE26Contr-R, GCGGTCCTCGGAGGCATCGAGT

^aIntroduced restriction sites are underlined. wt, wild type.