Efficient strategy for constructing duck enteritis virus-based live attenuated vaccine against homologous and heterologous H5N1 avian influenza virus and duck enteritis virus infection

Table 1 Primers used for generating pBAC-C-KCE, donor plasmid pRThGA and identification of the pBAC-C-KCE-HA

Purpose and primer	Sequence (5’ → 3’)	Sequence designation, restriction enzyme site and introduction sequence
BAC insertion^a
UL26-F	aaagtcgac ataacttcgtatagcatacattatacgaagttatgccgtatgaatgcgctgac	Sal I site (bold), Lox p sequence (italic)
UL26-R	ttaattaacgcggacaaaacgacgattac	Pac I site (bold)
gB-F	gtaatcgtcgttttgtccgcgttaattaatgaaaaagacggcggtacaat	Pac I site (bold)
gB-R	aagaatgcattcggcctgg
Red-F	ccaggccgaatgcattcttcgtggggtgtggtgcttttggt
Red-R	tcgagcggccgc tagggataacagggtaatccccaccttatatattctttcccaccct	Not I site (bold), I-isce I sequence (italic)
Amp-F	aaattaattaaggggataacgcaggaaagaac	Pac I site (bold)
Amp-R	aaattaattaaacgtcaggtggcacttttcg	Pac I site (bold)
Modification pRThGA^b
pCA-I-SceI-H1-F	aaatagggataacagggtaat gttgagcctttttgtggagtgggttaaattgtactagcgcgtttcgctttgcagtacatctacgtattagtcatcgctatta	I-isce I sequence (bold), Homology arm H1 (italic)
pCA-I-SceI-H2-R	aaatagggataacagggtaat tagcatgcataacttcgtataatgtatgctatacgaagttatgcggccgc cacacaggaaacagctatgaccatgattac	I-isce I sequence (bold), Homology arm H2 (italic)
Amp t-I-SceI-F	aaaattaccctgttatccctacacgttaagggattttggtcat	I-isce I sequence (bold)
OriT-R6K-I-SceI-R	aaaattaccctgttatcccta	I-isce I sequence (bold)
Identification HA^c
BAC-F	gagaacagaaaagaaagcgcgt
BAC-R	cgcagccacagaaaagaaacga

^aPrimers used for the construction of the BAC insertion vector. The restriction are marked in italics. ^bPrimers used for modification of donor plasmid pRThGA. ^cPrimers used for verification HA gene insertion into pBAC-C-KCE base on MAGIC.

ISSN: 1297-9716