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

Table 2 Primers used for SDM and cloning of the MHV spike and IBV nucleocapsid gene

Primer	Sequence (5′ ➔ 3′)	Function
XhoI-t-MHV-S.F01	CTCGAG TTATATTGGTGATTTTAGATGTATCCAG	Cloning MHV S
MHV-S-StyI-XhoI.R01	CTCGAGCCAAGGCCATTTCACATACATTTC	Cloning MHV S
p-MHV-S-SDM414.F02	AGCTTGTGAACTCAAACGGTG	SDM EcoRI
p-MHV-S-SDM414.R02	GGATACATCTAAAATCACCAATATAAC	SDM EcoRI
p-MHV-S-SDM2823.F03	TTACTATAAGTTCGAGACTGCC	SDM XhoI
p-MHV-S-SDM2823.R03	CACCCTGCATTAATGCAC	SDM XhoI
IBV-H52_N_ATG_FW	ACCATGGCGAGCGGTAAGA	N-transcript
IBV-M41-#2-IR-RV	TTTTTTTTTTTTTTTTTTTGCTCTAACTCTATACTAG	N-transcript

XhoI restriction enzyme sites in sequences of primers XhoI-t-MHV-S.F01 and MHV-S-StyI-XhoI.R01 are in italics; an additional thymidine residue to keep the MHV spike gene ectodomain sequence in frame with the IBV spike gene signal sequence is in bold. SDM = site directed mutagenesis