江西农业大学学报2013，Vol.35Issue(3)：620-625,6.
小麦淀粉分支酶基因SBE-Ⅱb植物表达载体的构建及表达鉴定
Construction of Plant Transformation Vector of Wheat Starch Branching Enzyme SBE-Ⅱb and Its Expression Identification
摘要
Abstract
The total RNA was isolated from wheat (Triticun aestivum L.) endosperm 15 days after pollination.One fragment of 2 511 bp of SBE Ⅱb gene was reversely transcribed and amplified from its mRNA using primer pairs designed according to the known SBE Ⅱb gene sequences (AY740401.1) in GeneBank,and the results of sequence analysis showed that the nucleotide sequence was the same with the known.The plant expression vector for pPZP35S was constructed.The plasmid was transformed into tobacco by leaf dish transformation,and four transgenic plants were obtained by kanamycin selecting and PCR analysis,and the results demonstrated that SBE Ⅱ b gene was transformed into tobacco cells,successfully.The differences between the wild plants and the transformed plants were obvious in the original plant morphology,and the transformed plants leaves turned dark and the plants were smaller,and SqRT-PCR and qRT-PCR analysis results indicated that the SBE Ⅱ b gene was expressed in the leaves of all the transformed plants.Compared with the check plants,the transcriptional level of the transformed plants changed significantly,however,the starch branching enzyme activity showed no difference between them.关键词
小麦/转基因/实时荧光定量(qRT-PCR)/淀粉分支酶基因(SBE-Ⅱb)Key words
wheat (Triticun aestivum L.) / transgene / quantitative real-time PCR (qRT-PCR) / starch branching enzyme gene (SBE Ⅱ b)分类
农业科技引用本文复制引用
王自布,齐军仓,李卫华,曹连莆,石培春..小麦淀粉分支酶基因SBE-Ⅱb植物表达载体的构建及表达鉴定[J].江西农业大学学报,2013,35(3):620-625,6.基金项目
教育部生物资源科学专业综合改革试点项目(2012287)、贵州省重点支持学科建设项目(2011231)和贵州师范学院博士启动基金项目(12BS028) （2012287）
