摘要
以小偃6号基因组DNA为模板,利用聚合酶链式反应扩增到1kb左右的核酸序列,将其克隆到pMD18-T载体上。经测序鉴定及序列分析后得知,该片段为小麦高分子量谷蛋白基因(HMW-GS1Dx2)上游启动子及信号肽编码区,共1050bp。将其与β-葡糖苷酸酶(GUS)基因编码序列融合,用基因枪法将构建的嵌合基因转入小麦未成熟种子及叶片,检测其瞬时表达活性。组织化学检测表明,由该启动子驱动的GUS融合基因能在种子中表达,而在叶片中却未见表达,从而证实此HMW-GS1Dx2启动子具有在小麦胚乳中高效表达的活性,为小麦胚乳生物反应器的研究奠定了基础。
To study the expression of high molecular weight glutenin gene promoter of Triticum aestivum in seeds,a fragment about 1 kb was amplified from wheat genome of Xiaoyan 6 by polymerase chain reaction (PCR) and inserted into pMD18 T vector.Sequencing and similarity analysis showed that the cloned fragment contained 1 050 bp nucleotides including HMW GS 1Dx2 promoter and signal peptide coding region.Reporter gene GUS (β glucuronidase) was fused to the fragment cloned,and introduced into wheat leaves and immature grain via particle bombardment.Gus expression was observed in the immature seeds of Triticum aestivum ,but not in the leaves.These results indicated that this high molecular weight glutenin gene promoter had the high endosperm specific expression activity.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2005年第5期95-99,共5页
Journal of Northwest A&F University(Natural Science Edition)
基金
国家转基因研究与产业化开发专项(JY03A-11-01)
陕西省农业分子生物学重点实验室项目
