Brief report of the construction of infectious DNA clones of South African genetic variants of grapevine virus A and grapevine virus B

Table 1 Oligonucleotide primers used in the construction of DNA clones of GVA and GVB genetic variants P163-M5 and 953-1, respectively, shown in Fig. 2

Virus	Primer code	Sequence (5′–3′)	Amplified fragment^a
GVA P163-M5	5′Ca35S.F1	GGATCCCATGGAGTCAAAGATTC
	3′Ca35S-5′M5.R	GGGAATCAAGTTAAATATTCGAGTCCCCCGTGTTCTCTCC	I
	3′Ca35S-5′M5.F	GGAGAGAACACGGGGGACTCGAATATTTAACTTGATTCC
	M5.573. Aat.R	GCATTGGTGCTTATCCTC	II
	M5.336.Aat.F	CACATGCTCTATAGAGTTGCACC
	M5.3512.Spe.R	CTTGACCTGGTTAGAGCGA	IV
	M5.2743.Spe.F	AGGCTGACAGACTGGCGCGGAGC
	M5.5403.Avr.R	CACTCAACGTAGGACAAGGACTC	V
	M5.4945.Avr.F	ATGGCAAAGAGGAACGG
	M5.T(30).R	CATATGT(30)GTCATAGTATGACAACCTAGC	VI
GVB 953-1	5′Ca35S.F2	ATGCGGCCGCCATGGAGTCAAAGATTC
	3′Ca35S-5′953.R	CGGAAGATTAAAGAAAAAGVGAACTTTATTGAGTCCCCCGTGTTCTCTCC	I
	3′Ca35S-5′953.F	GGAGAGAACACGGGGGACTCAATAAAGTTCGCTTTTTCTTTAATCTTCCG
	953.880.Age.R	ATGTGCTCCCTCTGGTGGCC	II
	953.2704.Avr.R	TGGCTCTTTGCTCGCACACC	IV^b
	953.2461.Avr.F	AGGGGTGGTATGACACTTGG
	953.5027.Xho.R	CTAACCAGATCCTGCTGAGC	V
	953.4838.Xho.F	CGAGGATGAGCCCCTATGGC
	953.T(35).R	AGTTCGACT(35)TCGTGTTTATTCACGCTTCTTTCACC	VI

In bold are shown parts of the sequence of the CaMV 35S promoter in various orientations
^aSee Fig. 2. Fragments III of GVA P163-M5 and GVB 953-1 were PCR amplified using fragments I and II as templates, and primers pair 5′Ca35S.F1/M5.573.Aat.R and 5′Ca35S.F2/953.880.Age.R, respectively, according to the method described by Peremyslov and Dolja (2007)
^bFragment IV of GVB 953-1 was amplified using primers pair 3′Ca35S-5′953.F/953.2704.Avr.R