Translational Health Science and Technology Institute (THSTI), Faridabad, India.
Animal Biotechnology Centre, National Dairy Research Institute (NDRI), Karnal, 132001, India.
Sci Rep. 2021 Dec 1;11(1):23193. doi: 10.1038/s41598-021-02551-1.
Tosyl activated magnetic beads were used for aptamer selection against PAG- 7 and 18 proteins of bovine origin. PAG proteins were immobilized on beads with further addition of biotin tagged aptamer library. The recognition of aptamers with PAG was identified by ST-HRP based approach which was colorimetric in nature. The selected aptamers were sequenced and at the same time several new aptamers were identified. Later M-fold structure and G-quadruplex score of aptamers were analyzed for their selection. Those aptamers having high G value and complex structure were chosen. In dot blot assay, aptamers recognized PAG protein in an animal after 42 days of artificial insemination which later given birth to a healthy calf. Further the cross reactivity with serum of 0th day animal (post AI) or with non pregnant animal serum was minimal. Aptamers have also shown interaction with PAG protein of buffalo origin. These selected aptamers have commercial application especially in development of biosensors for early detection of pregnancy in bovine.
对来源于牛的 PAG-7 和 18 蛋白，使用对甲苯磺酰基活化的磁珠进行适体筛选。将 PAG 蛋白固定在珠上，进一步添加带有生物素标记的适体文库。通过基于 ST-HRP 的方法识别与 PAG 结合的适体，该方法本质上是比色的。对所选适体进行测序，同时鉴定出了几个新的适体。随后，对适体的 M-fold 结构和 G-四链体评分进行了分析，以进行选择。选择那些具有高 G 值和复杂结构的适体。在点印迹分析中，在人工授精后 42 天的动物中，适体识别 PAG 蛋白，随后该动物产下了一只健康的小牛。此外，与 0 天动物（授精后）或未怀孕动物血清的交叉反应性最小。适体也显示出与水牛来源的 PAG 蛋白的相互作用。这些选定的适体具有商业应用，特别是在开发用于牛早期妊娠检测的生物传感器方面。
