Background

Development of genetically modified crops has rapidly increased in last few years. The most widely grown GM crops express genes that confer herbicide tolerance and insect resistance. Detection system of GM crops is important for safety evaluation before its consumption.

Objective

The purpose of this research is to detect GM crops, especially PAT, in food-samples.

Results

The bar gene (PAT protein, herbicide resistant) was cloned in pGEX-4T-1 and expressed by E. coli. The high-affinity PAT-specific single-stranded DNA (ssDNA) aptamers were obtained from a random DNA library. MOE docking study was performed to identify the potential binding region of the selected aptamers on PAT. Aptamer-linked immobilized sorbent assay (ALISA) method was used to detect PAT.

Conclusion

We screened aptamer against PAT for developing an efficient detection method. The selected PAT specific aptamers, HRPA-05 and HRPA-07, showed the distinct target binding behaviors, and detected PAT protein by aptamer-linked immobilized sorbent assay method with high efficiency and selectivity.

中文翻译：

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适体连接的固定吸附剂测定法，用于检测GMO标记，膦丝菌素乙酰转移酶（PAT）

背景

近年来，转基因作物的发展迅速增长。生长最广泛的转基因作物表达赋予除草剂耐受性和昆虫抗性的基因。转基因作物的检测系统对于食用前的安全性评估非常重要。

目的

这项研究的目的是检测食品样本中的转基因作物，尤其是PAT。

结果

所述b AR基因（PAT蛋白，除草剂抗性）中的pGEX-4T-1克隆并通过表达大肠杆菌。高亲和力的PAT特异性单链DNA（ssDNA）适体是从随机DNA库中获得的。进行了MOE对接研究，以鉴定PAT上所选适体的潜在结合区域。用适体连接的固定化吸附测定（ALISA）方法检测PAT。

结论

我们筛选了针对PAT的适体，以开发一种有效的检测方法。所选择的PAT特异性适体HRPA-05和HRPA-07显示出独特的靶结合行为，并且通过适体连接的固定吸附剂测定方法高效且选择性地检测到PAT蛋白。