Identification of a conserved linear B-cell epitope in the M protein of porcine epidemic diarrhea virus

Zhang, Zhibang; Chen, Jianfei; Shi, Hongyan; Chen, Xiaojin; Shi, Da; Feng, Li; Yang, Bin

doi:10.1186/1743-422X-9-225

Virology Journal

Table 1 Sequences of the primers used in this study

From: Identification of a conserved linear B-cell epitope in the M protein of porcine epidemic diarrhea virus

Peptides	Primer sequences (5’→3’)	Position	Size of amplicon
tM-His₆	F: ATACATATGAGCATTCGGTTGTGGCG	295–678	384bp
	R: CGCCTCGAGGACTAAATGAAGCACT
	F: TAAGGATCCAGCATTCGGTTGTGGCGCAG
GST-tM	R: CGGCTCGAGGACTAAATGAAGCACTTTCTCAC	295–678	384bp
GST-tM	F: TATCATATGAGCATTCGGTTGTGG	295–678	384bp
M1	R: TATCTCGAGTACGCCAGTAGCAAC	295–480	186bp
M1	F: GCGCATATGCTCACTACTTCTGTGAT	295–480	186bp
M2	R: GCGCTCGAGATAGACAATTGTTGTAG	358–540	183bp
M2	F: GCACATATGGTACAGGTAAGTCAAT	358–540	183bp
M3	R: CTACTCGAGGACTAAATGAAGCACT	478–678	201bp
M3	F: gatccGTACAGGTAAGTCAATTACCTAATTTCGTCACAGTCGCCA	478–678	201bp
M4	AGGCCACTACAACAATTGTCTATGGACGTGTTtaa c	478-549	72bp
	R: tcgag ttaAACACGTCCATAGACAATTGTTGTAGTGGCCTTGGCGA
	CTGTGACGAAATTAGGTAATTGACTTACCTGTACg
	F: gatccGGACGTGTTGGTCGTTCAGTCAATGCTTCATCTGGCACTG
M5	GTTGGGCTTTCTATGTCCGGtaa c	541–600	60bp
	R: tcgag ttaCCGGACATAGAAAGCCCAACCAGTGCCAGATGAAGCA
	TTGACTGAACGACCAACACGTCCg
M6	F: gatccTCAGCTGTGAGTAATCCGAGTGCGGTTCTCACAGATtaa c	592–627	36bp
M6	R: tcgag ttaCACAGCTGAGTAGTCGCCGTGTTTTGACCGGACATAg	592–627	36bp
M7	F: gatccTCAGCTGTGAGTAATCCGAGTGCGGTTCTCACAGATtaa c	619–654	36bp
M7	R: tcgag ttaATCTGTGAGAACCGCACTCGGATTACTCACAGCTGAg F: gatccCTCACAGATAGTGAGAAAGTGCTTCATTTAGTCtaa c	619–654	36bp
M8	R: tcgag ttaGACTAAATGAAGCACTTTCTCACTATCTGTGAGg	646–678	33bp
M9	F: gatccGGACGTGTTGGTCGTTCAGTCAATtaa c	541–564	24bp
M9	R: tcgag ttaATTGACTGAACGACCAACACGTCCg	541–564	24bp
M10	F: gatccCGTTCAGTCAATGCTTCATCTGGCtaa c	553–576	24bp
M10	R: tcgag ttaATTGACTGAACGACCAACACGTCCg	553–576	24bp
M11	F: gatccGCTTCATCTGGCACTGGTTGGGCTtaa c	565–588	24bp
M11	R: tcgag ttaAGCCCAACCAGTGCCAGATGAAGCg	565–588	24bp
M12	F: gatccACTGGTTGGGCTTTCTATGTCCGGtaa c	577–600	24bp
M12	R: tcgag ttaCCGGACATAGAAAGCCCAACCAGTg	577–600	24bp
M13	F: gatccGGTTGGGCTTTCTATGTCCGGtaa c	580–600	21bp
M13	R: tcgag ttaCCGGACATAGAAAGCCCAACCg	580–600	21bp
M14	F: gatccTGGGCTTTCTATGTCCGGtaa c	583–600	18bp
	R: tcgag ttaCCGGACATAGAAAGCCCAg
	F003A gatccGCTTTCTATGTCCGGtaa c
M15	R: tcgag ttaCCGGACATAGAAAGCg	586–600	15bp
M15	R: tcgag ttaGACATAGAAAGCCCAg	586–600	15bp
M16	F: gatccTGGGCTTTCTATtaa c	583–597	15bp
M16	R: tcgag ttaGACATAGAAAGCCCAg	583–597	15bp
M17	F: gatccTGGGCTTTCTATtaa c	583–594	12bp
M17	R: tcgag ttaATAGAAAGCCCAg	583–594	12bp

The introduced restriction enzyme sites (NdeI and XhoI or BamHI and XhoI) are underlined. At the 5’ and 3’ terminal of each forward strand there is a sequence “gatcc” and “taac” (in italics), respectively. At the 3’ and 5’ terminal of each reverse strand there is a sequence “g” and “tcgagtta” (in italics), respectively. When the forward and reverse oligonucleotides annealed they form a cohesive BamHI site or NdeI at the 5’ terminus and a cohesive XhoI site at the 3’ terminus. The bases “taa” and “tta” (in bold) were introduced into each pair of oligonucleotides to form a stop codon.

Back to article page

ISSN: 1743-422X

Contact us

Submission enquiries: journalsubmissions@springernature.com