Construction of a High-Quality Yeast One/Two-Hybrid cDNA Library from Cassava (Manihot Esculenta Crantz)

摘要

Cassava is an important food and potential biofuel crop with starch-rich edible tuberous roots. It is tolerant to multiple abiotic stresses and can produce an acceptable yield under the adverse climatic and nutrient-poor soil. However, the molecular mechanisms of starch accumulation and the abiotic stress response involved in cassava are less known. In the present study, total RNA was isolated from the mixed leaves, stems, flowers, fruits and storage roots of cassava, and a good quality cDNA library was constructed. The titers of the resultant cDNA library were ～1.15×10~7 pfu/ml and an average insert size of ～1.5 kb. Therefore, the cDNA library constructed in this study was highly efficient. It can be used in the yeast one/two - hybrid systems to investigate both known and new interactions between DNA sequence of interest and transcription factors involved in the metabolic pathways and signaling transduction pathways of cassava.