Computational protocol: Overexpression of the kiwifruit SVP3 gene affects reproductive development and suppresses anthocyanin biosynthesis in petals, but has no effect on vegetative growth, dormancy, or flowering time

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Protocol publication

[…] The A. chinensis SVP3 coding sequence was cloned under the control of the Cauliflower mosaic virus (CaMV) 35S promoter (35S:SVP3) into the pART277 binary vector () as previously described (). The resulting plasmid was transformed into Agrobacterium tumefaciens strain EHA105 for transformation into A. deliciosa and A. eriantha, and into GV3101 for transformation into N. tabacum. A construct with a reporter gene uidA (GUS; β-gluronidase) under the control of the CaMV 35S promoter (35S:GUS) in appropriate Agrobacterium strains was used to transform control plants (). Actinidia eriantha SVP3 (GenBank accession no. KJ123703) was amplified from the bud cDNA using gene-specific oligonucleotide primers, 5’-ATGGCGAGAGAGAAGATCAAGA-3’ (forward) and 5’-TGGTGTGACATTTCAAGTTCG-3’ (reverse). The predicted amino acid sequence alignment with A. deliciosa and A. chinensis SVP3 homologues was performed using Vector NTI version 9.0. (Invitrogen) Clustal W (opening 15, extension penalty 0.3). […]

Pipeline specifications

Software tools Vector NTI Advance, Clustal W
Application Protein sequence analysis
Organisms Actinidia deliciosa, Arabidopsis thaliana, Nicotiana tabacum
Chemicals Anthocyanins