Construction of RNAi Vector for LINS/LIN7 and INVINH Gene of Tomatoes Using Gateway Technology

摘要

Invertase inhibitor (INVINH) is a novel protein family in higher plants.It has become an important factor at the posttranslational level in the studies of plant photosynthetic operation sugar regulation of plant organ development and product quality.To allow a more precise comparison interaction between LIN5/LIN7 and INVINH gene of tornatoes,the paper constructed two RNAi vectors,in which the expression of LIN5/LIN7 and INV/NH was silenced using the Gateway technology.The interference fragment was cloned in entry clone via TOPO cloning.Entry vector pENTRTM/D-LIN5/LIN7 and pENTRTM/D-INVINH were recombination with expression vector PH7GWIWG2 (Ⅰ) using the LR clonase.Restriction enzymes of Xba Ⅰ,EcoR Ⅰ and HindⅢ were used to detect RNAi expression vector pH7GWIWG2 (Ⅰ)-LIN5/LIN7 and pH7GWIWG2 (Ⅰ)-INVINH to form the correct hairpin structure.The expression vector of RNAi was obtained successfully.