TABLE 1.

Primers used in this study.

Primes	Sequence (5′-3′)a	Application
XhoI-U	ATCACTCGAGAAAAGAATGGCCCAGGTGCAGCTGCAGG	pPICZαA-Nb6
XbaI-R	AGCTTCTAGATCAATGATGGTGGTGATGATGTGAGGAGAC
I588A-F	CCAAAGAAGACAACCGCCACAGATTCACCATCATT	PCR mutagenesis
I588A-R	AATGATGGTGAATCTGTGGCGGTTGTCTTCTTTGG
D590A-F	AAGACAACCATCACAGCATCACCATCATTCCTA
D590A-R	TAGGAATGATGGTGATGCTGTGATGGTTGTCTT
L643A-F	GACAGCTGTGCTTGTGCAGAGTATGATCCTGAAT
L643A-R	ATTCAGGATCATACTCTGCACAAGCACAGCTGTC
D646A-F	TTGTTTAGAGTATGCACCTGAATGGTTTGAA
D646A-R	TTCAAACCATTCAGGTGCATACTCTAAACAA
Y62A-F	TGGTGGTTCTACAACAGCCGCAGACTCCGTGAA
Y62A-R	TTCACGGAGTCTGCGGCTGTTGTAGAACCACCA
D64A-F	TTCTACAACATACGCAGCCTCCGTGAAGGGCCGATT
D64A-R	AATCGGCCCTTCACGGAGGCTGCGTATGTTGTAGAA
W105A-F	AGGGCGCGTACAGGCATGGCCTGTACTACGC
W105A-R	GCGTAGTACAGGCCATGCCTGTACGCGCCCT
P107A-F	CGCGTACAGTGGTGGGCTGTACTACGCGCGCTGAA
P107A-R	TTCAGCGCGCGTAGTACAGCCCACCACTGTACGCG
pBA-F	GCGAACTGACTGCCAAAGAAC
pBA-R	GCCCGGGGGAAAATGAAACCTC
ORF7-F	AGATCATCATCGCCCAACAAAAC	RT-qPCR
ORF7-R	GACACAATTGCCGCTCACTA
β-actin-F	TCCCTGGAGAAGAGCTACGA
β-actin-R	AGCACTGTGTTGGCGTACAG
F-6	GTATAGGTGTTGGCTCTATGC	(−) gRNA analysis
R-683	GGGAGCGGCAAGTTGGTTAACAC
F-12	GTGTTGGCTCTATGCCACGGC
R-343	TATAAAATAGACCCAGCACCC
(−) gRNA-actin-F	CTTCCTGGGCATGGAGTCC	β-actin analysis
(−) gRNA-actin-R	GGCGCGATGATCTTGATCTTC

^aMutated nucleotides are underlined. ^bRestriction sites are underlined and shown as italics.