Epitope mapping and characterization of a novel Nsp10-specific monoclonal antibody that differentiates genotype 2 PRRSV from genotype 1 PRRSV

Zhang, Zhibang; Wen, Xuexia; Dong, Jianguo; Ge, Xinna; Zhou, Lei; Yang, Hanchun; Guo, Xin

doi:10.1186/s12985-017-0782-9

Virology Journal

Table 1 Primers used for amplifying Nsp10 gene fragments and the corresponding amino acid sequence and size of each fragment

From: Epitope mapping and characterization of a novel Nsp10-specific monoclonal antibody that differentiates genotype 2 PRRSV from genotype 1 PRRSV

Name		Primer sequence (5′-3′)	Amino acid (aa) sequence	Size (aa)
A-1	F	GAATTCTGGGAAGAAGTCCAGAATG	¹GKKSR^……QTRRG¹⁰⁰	100
A-1	R	GGTACCTCATCCGCGGCGAGTCTGG	¹GKKSR^……QTRRG¹⁰⁰	100
A-2	F	GAATTCTTTAGTCTCCGTTAGGCG	¹⁰¹LVSVR^……TLQFP²⁰⁰	100
A-2	R	GGTACCTCAAGGGAATTGCAGCGTTG	¹⁰¹LVSVR^……TLQFP²⁰⁰	100
A-3	F	GAATTCTGCCCCCTCCCGTACCGG	²⁰¹APSRT^……MSMVN³⁰⁰	100
A-3	R	GGTACCTCAGTTGACCATGGACATAAG	²⁰¹APSRT^……MSMVN³⁰⁰	100
A-4	F	GAATTCTACGACCCGTGTGACCTAC	³⁰¹TTRVT^……CADLE⁴⁴¹	141
A-4	R	GGTACCTCATTCCAGGTCTGCGCAAATAG	³⁰¹TTRVT^……CADLE⁴⁴¹	141
B-1	F	GAATTCTGCCCCCTCCCGTACCGG	²⁰¹APSRT^……LRLLS²⁴⁰	40
B-1	R	GGTACCTCAACTGAGAAGCCTCAAG	²⁰¹APSRT^……LRLLS²⁴⁰	40
B-2	F	GAATTCTAATCACCTTGATGTCTTG	²³¹NHLDV^……DIMPQ²⁷⁰	40
B-2	R	GGTACCTCACTGAGGCATGATGTCA	²³¹NHLDV^……DIMPQ²⁷⁰	40
B-3	F	GAATTCTCATTGCTATGTATTTGAC	²⁶¹HCYVF^……MSMVN³⁰⁰	40
B-3	R	GGTACCTCAGTTGACCATGGACATAAG	²⁶¹HCYVF^……MSMVN³⁰⁰	40
C-1	F	GAATTCTCATTGCTATGTATTTGAC	²⁶¹HCYVF^……IWRFG²⁸⁰	20
C-1	R	GGTACCTCACCCGAACCTCCAGATGG	²⁶¹HCYVF^……IWRFG²⁸⁰	20
C-2	F	GAATTCTACCCAATTAAAGACCATC	²⁷¹TQLKT^……AIQPD²⁹⁰	20
C-2	R	GGTACCTCAATCTGGTTGAATGGCATC	²⁷¹TQLKT^……AIQPD²⁹⁰	20
C-3	F	GAATTCTCAGAATATCTGTGATGC	²⁸¹QNICD^……MSMVN³⁰⁰	20
C-3	R	GGTACCTCAGTTGACCATGGACATAAG	²⁸¹QNICD^……MSMVN³⁰⁰	20
D-1	F	GAATTCTCAGAATA…CAAACTTTGAGGTACC	²⁸¹QNICDAIQPDYRDKL²⁹⁵	15
D-1	R	GGTACCTCAAAGTTTG…TATTCTGAGAATTC	²⁸¹QNICDAIQPDYRDKL²⁹⁵	15
D-2	F	GAATTCTGCCATTC…GGTCAACTGAGGTACC	²⁸⁶AIQPDYRDKLMSMVN³⁰⁰	15
D-2	R	GGTACCTCAGTTGACC…GAATGGCAGAATTC	²⁸⁶AIQPDYRDKLMSMVN³⁰⁰	15
E-1	F	GAATTCTGCC…GAT…GAC…CTTTGAGGTACC	²⁸⁶AIQPDYRDKL²⁹⁵	10
E-1	R	GGTACCTCAAAG…GTC…ATC…GGCAGAATTC	²⁸⁶AIQPDYRDKL²⁹⁵	10
E-1- mutant	F	GAATTCTGCC…TGT…GAG…CTTTGAGGTACC	²⁸⁶AIQPCYREKL²⁹⁵	10
E-1- mutant	R	GGTACCTCAAAG…CTC…ACA…GGCAGAATTC	²⁸⁶AIQPCYREKL²⁹⁵	10
E-2	F	GAATTCTGCCATTCAA…GACAAATGAGGTACC	²⁸⁶AIQPDYRDK²⁹⁴	9
E-2	R	GGTACCTCATTTGTC...TTGAATGGCAGAATTC	²⁸⁶AIQPDYRDK²⁹⁴	9
E-3	F	GAATTCTATTCAA…GACAAACTTTGAGGTACC	²⁸⁷IQPDYRDKL²⁹⁵	9
E-3	R	GGTACCTCAAAGTTTGTC...TTGAATAGAATTC	²⁸⁷IQPDYRDKL²⁹⁵	9

The forward (F) and reverse (R) primers contain EcoRI and kpnI recognition sites, respectively (underlined). The superscript numbers indicate the relative location of the amino acids in Nsp10 protein

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ISSN: 1743-422X

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