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甘蔗(Saccharum officinarum)是制造蔗糖、提炼乙醇的主要原料。随着甘蔗转基因育种的开展,建立一套快速精准转基因分子检测技术在甘蔗转基因研究中有重要的应用前景。本研究以二价抗虫转基因甘蔗为材料,利用多重PCR建立同时检测Bar基因、Bt基因和马铃薯蛋白酶抑制剂基因(PinⅡ基因)。研究表明,该多重PCR检测体系的最佳退火温度为53℃,有效扩增Bar基因(415 bp)、Bt基因(780 bp)和PinⅡ(1 000 bp)3个基因片段。多重PCR技术进行外源基因的检测,节省时间、降低成本、提高效率并可以对该转化植物进行特异性鉴定。
Saccharum officinarum is the main raw material for making sucrose and ethanol. With the development of sugarcane transgenic breeding, the establishment of a set of rapid and accurate transgenic molecular detection technology in sugarcane transgenic has important application prospects. In this study, bivalent insect-resistant transgenic sugarcane was used as material to establish simultaneous detection of Bar gene, Bt gene and potato proteinase inhibitor gene (PinⅡ gene) by multiplex PCR. The results showed that the optimum annealing temperature of this multiplex PCR system was 53 ℃, which effectively amplified Bar gene (415 bp), Bt gene (780 bp) and Pin Ⅱ (1 000 bp). Multiplex PCR allows for the detection of exogenous genes, saving time, reducing costs, increasing efficiency and allowing specific identification of the transformed plant.