TABLE 3.
Oligonucleotides used for targeted mutagenesis and qPCR amplificationa
| Source | Nameb | Sequence, 5′–3′c |
|---|---|---|
| E. coli K-12 MC1061 | ||
| pACY177 | Km_Fw | TCTCTGATGTTACATTGCAC |
| Km_Rv | CAGCGTAATGCTCTGC | |
| LKm_Rv | GATTGTCGCACCTGATTGCC | |
| E. coli K-12 MA8 | ||
| pKNOCK-Gm | pKNOCK_Fw | CATAAGCCTGTTCGGT |
| pKNOCK_Rv | CGGGGGATCCACTAG | |
| Rhodospirillum rubrum S1H | ||
| Genomic DNA | ccrHR1_Fw | AGAACTAGTGGATCCGGCGTTCTTTGACAAGACCA |
| ccrHR1_Rv | AATGTAACATCAGAGAGTTGAAACCTCTAGGCCGCT | |
| ccrHR2_Fw | GCAGAGCATTACGCTGAAGAGGTGCGCAAGTTCG | |
| ccrHR2_Rv | ACCGAACAGGCTTATGTCGCTCAGGTGCCATTC | |
| LKm_Fw | TATCGGCGCCTTCGGTGTAT | |
| Ccr_Fw | GTCAACTACAACGGGATCTGGG | |
| Ccr_Rv | GAAATCGCGCTTGGTCTCATC | |
| Glts_Fw | GATCGGCTATTGCAACTGCG | |
| Glts_Rv | CAGATCCTCCATCGACTGGC |
a
All the oligonucleotides were designed using Primer3Plus v2.4.0 and checked for specificity using Snapgene Viewer software (GSL Biotech) and for the formation of dimers and secondary structures using the Multiple Primer Analyzer webtool (ThermoFisher Scientific) and were obtained from Eurogentec S.A.
b
Fw, forward primer; Rv, reverse primer.
c
Nucleotide sequences based on the sequences of the pACY117 plasmid (GenBank accession no. X06402.1), the pKNOCK-Gm plasmid, and Rhodospirillum rubrum strain ATCC 11170 (NCBI reference sequence; NC_007643.1). The underlined sequences are end-terminal homology sequences required for GeneArt Seamless cloning.