Development of a novel Newcastle disease virus (NDV) neutralization test based on recombinant NDV expressing enhanced green fluorescent protein

Table 2 Oligonucleotides used for cloning of the supporting plasmids, construction of pFLC-LS1-1eGFP, and confirmation of virus recovery

Primers	Sequences (5′ → 3′)^a,b	Restriction site	Nucleotide Position^c
Supporting plasmid primers
Nfor	GGAATTC GCCACCATGTCTTCCGTATTTGATG	EcoRI	116–140
Nrev	GCGGCCGCTCAATACCCCCAGTCGGTGT	NotI	1572–1591
Pfor	TGAATTC GCCACCATGGCCACCTTTACAGATGC	EcoRI	1886–1906
Prev	GCGGCCGCTTAGCCATTTAGAGCAAG	NotI	3057–3074
Lfor	TACTAGT GCCACCATGGCGAGCTCCGGTCCTGAA	SpeI	8380–8401
Lrev	GCGGCCGCTTAAGAGTCACAGTTACTGT	NotI	14,976–14,995
pFLC-LS1-1eGFP construction
NDV116_F	GCCAACATGTCTTCCGTATTTGATG	PciI	116–140
NDV3211_R	CGATCATTCAGTGGGGCTGAGG	BbvCI	3190–3211
Confirmation primers
M + 4100 ^d	AGTGATGTGCTCGGACCTTC	N.A.^e	4100–4119
NDV4394_R	GAGACGCAGCTTATTTCTTAAAAGG	N.A.	4370–4394
NDV14035_F	TCCAGGGTCCAATCAAAGCT	N.A.	14,035–14,054
NDV15003_R	GATTTTCGTTAAGAGTCACAGTTACTG	N.A.	14,977–15,003

^aRestriction sites sequences are underlined
^bKozak sequence is shown in bold
^cThe positions where primers bind are according to the published NDV sequence (GenBank Accession no. Y18898)
^dThis primer was taken from Wise et al. [26]
^eNot applicable

ISSN: 1743-422X