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. 2020 Nov 4;11:571807. doi: 10.3389/fmicb.2020.571807

TABLE 3.

Protein pair combinations performed in the BiFC heterologous assays.

Heterologous assays p29/MP CiLV-C2 N/MP CiLV-N P/N CiLV-N P/MP CiLV-N N/MP OFV
ORF-NYFP + ORF-CYFP + + +
ORF-NYFP + CYFP-ORF + +
NYFP-ORF + ORF-CYFP +
NYFP-ORF + CYFP-ORF + + +
ORF-CYFP + ORF-NYFP + + +
ORF-CYFP + NYFP-ORF + +
CYFP-ORF + ORF-NYFP + + +
CYFP-ORF + NYFP-ORF + + +
Negative controls
CYFP-ORF + NYFP-cyt
ORF-CYFP + NYFP-cyt
NYFP-ORF + CYFP-ER

P/N OFV P/MP OFV p29 CiLV-C/p29 CiLV-C2 MP CiLV-C/MP CiLV-C2 p29 CiLV-C/MP CiLV-C2

ORF-NYFP + ORF-CYFP + +
ORF-NYFP + CYFP-ORF + + +
NYFP-ORF + ORF-CYFP + +
NYFP-ORF + CYFP-ORF + + +
ORF-CYFP + ORF-NYFP + + + +
ORF-CYFP + NYFP-ORF + + +
CYFP-ORF + ORF-NYFP + + + +
CYFP-ORF + NYFP-ORF + + +
Negative controls
CYFP-ORF + NYFP-cyt
ORF-CYFP + NYFP-cyt
NYFP-ORF + CYFP-ER

p29 CiLV-C2/MP CiLV-C N CiLV-N/N OFV MP CiLV-N/MP OFV N CiLV-N/MP OFV N OFV/MP CiLV-N

ORF-NYFP + ORF-CYFP +
ORF-NYFP + CYFP-ORF + + +
NYFP-ORF + ORF-CYFP +
NYFP-ORF + CYFP-ORF +
ORF-CYFP + ORF-NYFP + +
ORF-CYFP + NYFP-ORF + +
CYFP-ORF + ORF-NYFP + + +
CYFP-ORF + NYFP-ORF + +
Negative controls
CYFP-ORF + NYFP-cyt
ORF-CYFP + NYFP-cyt
NYFP-ORF + CYFP-ER

Intra-association control

MP CiLV-C2/MP OFV p29 CiLV-C2/N OFV p29 CiLV-C2/MP OFV MP CiLV-C2/N OFV MP CiLV-C/p24 CiLV-C

ORF-NYFP + ORF-CYFP + +
ORF-NYFP + CYFP-ORF +
NYFP-ORF + ORF-CYFP +
NYFP-ORF + CYFP-ORF +
ORF-CYFP + ORF-NYFP + +
ORF-CYFP + NYFP-ORF + +
CYFP-ORF + ORF-NYFP + + +
CYFP-ORF + NYFP-ORF +
Negative controls
CYFP-ORF + NYFP-cyt
ORF-CYFP + NYFP-cyt
NYFP-ORF + CYFP-ER

Inter-association controls

CP AMV/MP CiLV-C N TSWV/MP CiLV-C

ORF-NYFP + ORF-CYFP
ORF-NYFP + CYFP-ORF
NYFP-ORF + ORF-CYFP
NYFP-ORF + CYFP-ORF
ORF-CYFP + ORF-NYFP n.d
ORF-CYFP + NYFP-ORF n.d
CYFP-ORF + ORF-NYFP n.d
CYFP-ORF + NYFP-ORF n.d
Negative controls
CYFP-ORF + NYFP-cyt
ORF-CYFP + NYFP-cyt
NYFP-ORF + CYFP-ER

The symbols − and + correspond to the absence and presence of fluorescence signals, respectively. NYFP and CYFP correspond to half N- and C-terminal of the YFP protein, respectively. OFV = OFV-citrus. n.d = non-determined. Additional negative controls are presented for intra- and inter-association assays.