Background
Materials and methods
Yeast two-hybrid (Y2H) screen
Primer | Sequence(5′-3′) | Usage |
---|---|---|
BD-C-F | CCGCATATGTCCGACACAAATGCAGAAGGGGC | Amplification of C Protein |
BD-C-R | GCGGAATTCTTATCCCACTGCAACCTGAA | Amplification of C Protein |
HA-C-F | CGGAATTCGCTCCGACACAAATGCAGAA | Amplification of C Protein |
HA-C-R | GGGGTACCTTATCCCACTGCAACCTGAAAC | Amplification of C Protein |
Myc-PIAS4-F | TATCTCGAGCGATGGCGGCGGAACT | Amplification of PIAS4 |
Myc-PIAS4-R | TATGGATCCGCAGGCCGACACCAGA | Amplification of PIAS4 |
AD-PIAS4-F | TTCATATGGCGGCGGAACTGGTGGAG | Amplification of PIAS4 |
AD-PIAS4-R | ATACTCGAGTCAGCAGGCCGACACCAGAC | Amplification of PIAS4 |
ISG15-F | GCAGACCAGTTCTGGCTGTCT | Amplification of ISG15 |
ISG15-R | CCAGCGGGTGCTCATCAT | Amplification of ISG15 |
MX1-F | GAGGTGGACCCCCAAGGA | Amplification of MX1 |
MX1-R | CCACCAGATCGGGCTTTGT | Amplification of MX1 |
OAS1-F | CCAAGTCAAACAAGCCATCGA | Amplification of OAS1 |
OAS1-R | CACATCGGAAACACCTCTCCTT | Amplification of OAS1 |
IFITM3-F | CGTGTGGTCCCTGTTCAAC | Amplification of IFITM3 |
IFITM3-R | CCATCTTCCGGTCCCTAGAC | Amplification of IFITM3 |
GAPDH-F | AAGGCCATCACCATCTTCCA | Amplification of GAPDH |
GAPDH-R | CCACCACATACTCAGCACCAGCAT | Amplification of GAPDH |
Plasmids
Cell, viruses and virus titration assays
Transfection of plasmid DNA
Virus infection
Co-immunoprecipitation (co-IP)
Confocal imaging
RNA interference
Western blot analysis
qRT-PCR
Statistical analysis
Results
Identification of bovine PIAS4 as an interacting partner of BVDV C protein
Prey | No. of Genbank accession | No. of hits | similarity |
---|---|---|---|
Bos taurus small glutamine-rich tetratricopeptide repeat (TPR)-containing, alpha (SGTA) | NM_001038030 | 15 | 89% |
Bos taurus protein inhibitor of activated STAT, 4 | BC123606 | 13 | 93% |
Bovine natural resistance-associated macrophage protein (Nramp) | NM_174652 | 5 | 92% |
Bos taurus calcium modulating ligand (CAMLG) | NM_001037625 | 6 | 93% |
Bos taurus cysteine-rich with EGF-like domains 1 (CRELD1) | NM_001014851 | 10 | 92% |
Bos taurus transmembrane emp24-like trafficking protein (TMED10) | NM_001040549.2 | 4 | 92% |
Bos taurus ribosomal protein L6 (RPL6) | BT021544.1 | 7 | 90% |
Bos taurus tetratricopeptide repeat domain (TTC1) | NM_001034217.2 | 3 | 93% |
Bos taurus reticulon-4 (RNT4), transcript variant 1 | NM_001075138.1 | 5 | 93% |
Bos taurus hypothetical protein LOC617407 | BC108156.1 | 10 | 88% |
Predicted:Ovis aries vascular cell adhesion protein 1-like,transcript variant 3 | XM_004002235.1 | 5 | 89% |
Bos taurus eukaryotic translation initiation factor 3, subunit E (EIF3E) | NM_001034603.1 | 2 | 80% |
Bos taurus protein-L-isoaspartate (D-aspartate) O-methyltransferase | BC109663.1 | 2 | 93% |