Table 1.
Table 1 A. PCR primers and annealing temperatures for trans-sialidase and TcPKAc genes | ||
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Trans-sialidase genes and TcPKAc gene |
PCR primers | Annealing Temperature |
Tc00.1047053508563.20 | Forward- (Nhe1) CTAGCTAGC ATGCTCTCACGTGTTGCTGC Reverse- (Xbal1) GATCTCTAGA TCACGCAGTCGCAAAGCCCC |
55°C |
Tc00.104753509187.10 | Forward- (Nhe1) CTAGCTAGCATGTCCCGGCGTGTGTTTACT Reverse- (Xbal1) GATCTCTAGATCACGCAGCCGCAAACCC |
55°C |
Tc00.1047053507839.40 | Forward- (Xho1) CCGCTCGAG ATGCTCTCACGTGTTGCTG Reverse- (Sal1) ACGCGTCGAC TCACGCAGCCGCAAAGCCC |
66°C |
Tc00.104753510111.20 | Forward- (Xho1) CCGCTCGAG ATGCTCTCACGTGTTGCTG Reverse- (Sal1) ACGCGTCGAC TCACGCAGTCGCAAGGCCC |
66°C |
TcPKAc (Genbank, AY055783) | Forward- (EcoR1) CGGAATTCATGTCAGGGGAGGCAAATTATG Reverse- (Pst1) AACTGCAGTTAGAACCCAATAAACTCCGCCTG |
55°C |
Table 1B. In silico analysis of PKA phosphorylation site. | |
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Each of trans-sialidase protein sequences was analyzed against PROSITE patterns and profiles (http://us.expasy.org/prosite). All of these proteins contain typical PKA phosphorylation sites. The numbers indicate the location of the sites in the various protein sequences. | |
Trans-sialidase genes | cAMP- dependent protein kinase phosphorylation site |
Tc00.1047053508563.20 | 113 – 116: KKgT; 120 – 123: RRdS; 465 – 468: RRvS; 624 – 627: KKlT |
Tc00.104753509187.10 | 17 – 20: RRvT; 635 – 638: KKiS |
Tc00.1047053507839.40 | 17 – 20: RRvT; 239 – 242: KRiS; 633 – 636: KKaS |
Tc00.104753510111.20 | 17 – 20: RRvT; 468 – 471: RRgS; 649 – 652: KKrS; 699 – 702: KKpS |
Tc00.1047053508285.60(SA1.1) | 17 – 20: RRvT; 119 – 122: RRdS; 466 – 469: RRdS; 624 – 627: KKpT |
TcTS 6 11/2 | 12 – 15: KRqS |
Table 1C. Myristoylation sites exist in TcPKAc | |
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The PROSITE program was used to identify N-myristoylation sequences in both TcPKAc isoforms | |
TcPKAc | Myristoylation site |
Protein kinase A catalytic subunit isoform-1 (Tc00.1047053508461.310) |
306 – 311: GLlqTD; 317 – 322: GTlkGG |
Protein kinase A catalytic subunit isoform- 2 (Tc00.1047053508461.280) |
25 – 30: GstlGA; 250 – 255: GLlqTD; 261 – 266: GTlkGG |