Table I.

Oligonucleotides used in this study for amplification or mutagenesis of HCoV sequences

Oligonucleotide	Oligonucleotide sequence (5′ to 3′)1-a	Nucleotide no.1-b	Polarity	Use
134	AAAAGGATCCAATGTGACATACACTGACCAGTATCTC	626–652	Forward	PCR
135	AAAACTGCAGTTACTGAAGAGCAATCACTCTCCAAACAGAG	1231–1258	Reverse	PCR
110	TATAATACGACTCACTATAGGGCACCATGAAACGTTGGCTTGGCATTTTGCTAG	2567–2591	Forward	PCR
111	TATAATACGACTCACTATAGGGCACCATGGTTATAGATGGAGGCAAAATATGGAAT	2441–2467	Forward	PCR
103	TGAGGAATTCTTATTGGTAGAAAGCTACATTGTCATG	4124–4147	Reverse	PCR
105	TCCCGAATTCTTACACTAAACCAGACACAAAATCCTT	4577–4600	Reverse	PCR
107	TTATGGTTTAGGTTTCACTGTATTAACAGG	5926–6022	Reverse	PCR
112	TTGGATAACAATGCATGGGTTAACTCAGTTATGTTACA	3440–3477	Forward	Codon mutagenesis
113	TGAGTTAACCCATGCATTGTTATCCAATTGTTTGAGTATC	3427–3466	Reverse	Codon mutagenesis
188	ATGTCATTTTCTGATGACATGGAAATGAAAGACATTGAAC	2990–3029	Forward	Codon mutagenesis
189	CATTTCCATGTCATCAGAAAATGACATTTTACCACCAGC	2978–3016	Reverse	Codon mutagenesis
139	TGGTAATGACCACAAGAGACCGCACCACGAAAAATTGATG	3873–3912	Reverse	PCR
165	TTTCGTGGTGCGGTCTCTTGTGGTCATTACCAGACT	3881–3916	Forward	PCR
210	ATATGGTCTCATGGTTGTTATCCAATTGTTTGAGTATCT	3426–3451	Reverse	Deletion mutagenesis
211	ATATGGTCTCAACCAAATACAATTAACTGGTATACTTGA	3476–3501	Forward	Deletion mutagenesis
212	TATAGGTCTCAGGCGTTGTTATCCAATTGTTTGAGTATC	3427–3451	Reverse	Codon mutagenesis
213	TATAGGTCTCACGCCTGGGTTAACTCAGTTATGTTACAA	3454–3478	Forward	Codon mutagenesis
214	TATAGGTCTCAATGCATTATTATCACTAGTTTTTAACAC	5369–5392	Reverse	Codon mutagenesis
215	TATAGGTCTCAGCATGGGTGAATGCTGTTTGTATTGCAC	5396–5420	Forward	Codon mutagenesis
167	ACCCGGTCTCAATTATTATCACTAGTTTTTAACACTCTAATACC	5360–5393	Reverse	Deletion mutagenesis
168	ACCCGGTCTCTTAATGCACTACAGTATTCGAAACCCCATTTTATTTC	426–5448	Forward	Deletion mutagenesis
137	TTTTTTCCATGGGCTACTTCCGTCTTATGGCCAGTCCAA	2006–2033	Forward	PCR
39	AATCAAAGAACTTCTGTGCC	6046–6065	Reverse	PCR

^1-aThe underlined sequences were required for cloning, mutagenesis, and expression of HCoV sequences. The boldface sequences present in oligonucleotides 110 and 111 indicate the T7 RNA polymerase promotor.

^1-bHCoV nucleotide numbers correspond to the sequence published by Herold et al. (47).