A reverse genetics system for avian coronavirus infectious bronchitis virus based on targeted RNA recombination

Table 1 Plasmids used for generation of the donor plasmids p-IBV and p-mIBV

Plasmid	Genes	Coordinates	Length (nt)	3′-end RES	Surrounding RES	Inserted at 3′-end of
p-IBV-5	T7	n.a.	18	n.a.	n.a.	n.a.
	GG	n.a.	2	n.a.	n.a.	n.a.
	IBV 5′-UTR	1–497	497	BstBI	n.a.	n.a.
p-IBV-1b	IBV 3′-end 1b, S (ss)	19,610–20,379	770	XhoI	BstBI	p-IBV-5
p-IBV-S	IBV S (ec)	20,379–23,590	3211	StyI	XhoI	p-IBV-5-1b
p-IBV-SIR	IBV S (tm, en), 3a, 3b, E, M	23,591–25,318	1728	EcoRI	StyI	p-IBV-5-1b-S
p-IBV-3 T	IBV 5a, 5b, N, 3′-UTR	25,319–27,630	2322	n.a.	EcoRI	p-IBV-5-1b-S-SIR
	PolyA tail	27,631–27,730	100	MssI, PacI	n.a.	n.a.
p-MHV-S	MHV S (ec)	n.a.	3757	StyI	XhoI	p-IBV-5-1b

IBV gene fragments were generated in pUC57-simple, the MHV spike ectodomain derived from pTUG was ligated into pJet1.2. Plasmid names and spike protein domain abbreviations refer to Fig. 1. (ss) = signal sequence; (ec) = ectodomain; (tm) = transmembrane domain; (en) = endodomain; n.a. = not applicable. RES = restriction enzyme site

ISSN: 1743-422X