Table 2 The sequences of primers used in this study
Target | Direction | Sequences |
|---|---|---|
For qPCR | ||
ASB3 | Forward | 5′-CATACTTATTTCATCGGGTGC-3′ |
Reverse | 5′-GGTAACTGCCAACTGTCCTC-3′ | |
GAPDH | Forward | 5′-CTCCTCCTGTTCGACAGTCAGC-3′ |
Reverse | 5′-CCCAATACGACCAAATCCGTT-3′ | |
For vector construction (italics indicates restriction enzyme recognition sequence) | ||
WT ASB3 | Forward | 5′-GCGCTCGAGATGGATTTTACAGAGGCT-3′ |
Reverse | 5′-GGCGCGGCCGCTTATCCATCTTGAATAGCTG-3′ | |
ΔSOCS | Forward | 5′-GCGCTCGAGATGGATTTTACAGAGGCT-3′ |
Reverse | 5′-GGGATCGATTTACCTTTCAACAGCTGGTG-3′ | |
For point mutation generation (capital letter indicates mutated nucleotide) | ||
G135E | Forward | 5′- ggctgttgcttcaacacgAagcaaatgttaatggatc-3′ |
Reverse | 5′-gatccattaacatttgctTcgtgttgaagcaacagcc-3′ | |
K339I | Forward | 5′-gtgaacattcttttgaTatatggagcccagata-3′ |
Reverse | 5′-tatctgggctccatatAtcaaaagaatgttcac-3′ | |
R362C | Forward | 5′-cgagaagttttcgatatttTgctactttttgaggaaagg-3′ |
Reverse | 5′-cctttcctcaaaaagtagcAaaatatcgaaaacttctcg-3′ | |