TABLE 3.
Primers used in this studya
| Target or primer | Sequence (5′–3′) | Source or reference |
|---|---|---|
| “Ca. Liberibacter asiaticus” confirmation | ||
| OI1 | GCGCGTATGCAATACGAGCGGC | 48 |
| OI2c | GCCTCGCGACTTCGCAACCCAT | 48 |
| CG03F | RGGGAAAGATTTTATTGGAG | 49 |
| CG05R | GAAAATAYCATCTCTGATATCGT | 49 |
| Quantitative RT-PCR | ||
| Lcr_gloA_F | GTTGTGTAGTCTTCAGTATCCCAG | This study |
| Lcr_gloA_R | GGGTCTCTATGAAGTTGATCGTC | This study |
| Lcr_gloB_F | CATGCGGATCATACAAAAGGC | This study |
| Lcr_gloB_R | CCTTCAGAAAGCACATGATCAAC | This study |
| CLas_gloB_F | GATTCAAACATTTCAGCATAACTATCTTC | This study |
| CLas_gloB_R | GATCATTTTCTTTGCGTCGGAG | This study |
| Lcr_TPI_F | CGGCAACATTAGTGTATTTGGC | This study |
| Lcr_TPI_R | ACAAAATCAGCTCCACAATCAAC | This study |
| CLas_TPI_F | CCGTACCAATCGCCCATATAG | This study |
| CLas_TPI_R | AGATTGTAGTTTGCCGAGTGAG | This study |
| Lcr_GAPDH_F | GTCTTATCAGCTCCTCCACAAG | This study |
| Lcr_GAPDH_R | GTGATCCAAAACAGCTTCCG | This study |
| CLas_GAPDH_F | AGTTGAGTCAAGACGCGATG | This study |
| CLas_GAPDH_R | CCTATGATTTTCACCTCTCCTGG | This study |
| CLas_nttA_F | CTACATTCCCGACCAATCCAA | This study |
| CLas_nttA_R | GGCGCTGTCATGATTAATCTTATG | This study |
| Lcr_gyrB_F | TCTTCACCAGCATCTCCAAC | This study |
| Lcr_gyrB_R | CACTTTCATCTTGGCTGCG | This study |
| CLas_gyrB_F | TTGAACAAGCTGTAATTTCTGG | 8 |
| CLas_gyrB_R | ATCTGTTTGCCAATTTAGAAGC | 8 |
| Lcr_prfA_F | AGGCTCAAGTTGGTTCAGG | This study |
| Lcr_prfA_R | ATATCACCCTCCAACATGCG | This study |
| CLas_prfA_F | TGTCTGAATCGCCTTCTGTC | 8 |
| CLas_prfA_R | GATCACCGATGACAGTATGC | 8 |
| Cloning in pUFR071 | ||
| Lcr_gloA_F | GGTACCAGGAGACATACTATGGTACGTGTTA | This study |
| Lcr_gloA_R | AAGCTTTTACCAGGTTCCTATGTTAGGCAT | This study |
| Xal_gloA_F | GAATTCAGGAGTTTCGCGATGAAATACCTGCATGCC | This study |
| Xal_gloA_R | AAGCTTTCACCATACTCCGGTATTGGGCA | This study |
| CLas_nttA_F | GAATTCAGGAGCGTTAAGATGTCGGAGGCGAAGA | This study |
| CLas_nttA_R | AAGCTTTTATTCTTTACTAATAAGCTGAGTATATTC | This study |
| Marker interruption | ||
| Lcr_gloA_internal_F | GCTCTTTGGGTTCTAGAGGATTAT | This study |
| Lcr_gloA_internal_R | CGTGTTAAAGACCTTGAGCATTC | This study |
| Lcr_lcrI_internal_F | CCATTGATCTGGTGCTGTTTATC | This study |
| Lcr_lcrI_internal_R | TTGGGCTTGGACGACTTTTGC | This study |
| Lcr_sut_internal_F | TATGCTTGGGAGCATGTTAGG | This study |
| Lcr_sut_internal_R | CAAGAACCGGAAGAGCGATAG | This study |
a
Abbreviations used in target and primer designations: gloA and gloB, glyoxalase I and II; TPI, triosephosphate isomerase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; gyrB, DNA gyrase subunit B; prfA, peptide chain release factor 1; nttA, ATP/ADP translocase; lcrI, restriction subunit R; sut, sugar transporter. In primer designations, CLas, Lcr, and Xal denote “Ca. Liberibacter asiaticus,” L. crescens, and X. albilineans, respectively, and “F” and “R” denote forward and reverse, respectively. Restriction sites for cloning are italicized in the corresponding sequences.