Constructing pBin61: The 35Spro-35Ster expression cassette of pJIT61 was excised as a kpnI-XhoI fragment and mobilised into the pBin19 binary vector T-DNA. The cassette was inserted into KpnI-SalI digested pBin19, therefore eliminating the SalI site. This led to plasmid pBin61. You will find the sequence of the vector as a MS Word file below. Inserting suppressors of gene silencing into pBin61: The SmaI site in pBin61 was used to clone all the viral suppressors in sense orientation. Blunt-end PCR-amplified fragments of the suppressors were used for cloning. It is therefore not possible to excise the suppressor sequence from the pBin61 vector. You will find sequences of each individual suppressor constructs cloned in pBin61 below.
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