花生高蛋白转化体T-DNA整合位点侧翼序列分析

Isolation of Peanut DNA Sequences Flanking T-DNA Insertion Sites from Transgenic Seeds with Elevated Protein Content

利用山东省花生研究所分子育种实验室改建的植物表达载体,采用自创的农杆菌直接注射法转化花生品种花育22号。运用近红外扫描技术分析单粒花生主要脂肪酸、含油量和蛋白质含量,获得了蛋白质含量明显提高的转化体。通过DNA walking技术获取T-DNA左边界侧翼序列3条、右边界侧翼序列1条。经Blast分析,在GenBank中找到有一定相似性的序列。这是花生上应用T-DNA标签法的首次尝试。

Plant expression vectors constructed at Molecular Breeding Laboratory of Shandong Peanut Research Institute were transferred into peanut cultivar Huayu 22 via an in planta protocol. NIRS was used to predict main quality attributes, including protein, oil, oleate and linoleate content, in the resultant single peanut seeds. Peanut transformants with elevated protein content were identified by NIRS and confirmed by Kjeldhal nitrogen determination procedure, and 4 peanut DNA sequences flanking T--DNA insertion sites were isolated. Blast analysis of these sequences revealed some similarity to the sequences deposited in GenBank. This represented the first attempt of T--DNA tagging in peanut.