Department of Life Science, Sharda University, Greater Noida, India.
Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana, India.
Mol Biol Rep. 2021 May;48(5):3925-3934. doi: 10.1007/s11033-021-06388-9. Epub 2021 May 20.
Studying the maternal oocyte-specific genes, in farm animals is a significant step towards delineating the underlying mechanisms that regulate oocyte quality, early embryonic development and survival. With the creation of buffalo oocyte-specific subtracted cDNA library, it has raised new questions which need to be answered. The present study has characterized one of the ESTs selected from the library and highlighted its importance in the oocyte quality. The selected EST was made full length by RLM-RACE and four transcript variants were identified. Bioinformatics analysis indicated the novelty of full-length transcript along with conserved intergenic nature. The largest transcript was identified as long intergenic noncoding RNA based upon coding potential calculator output. The expression analysis at different hours of oocyte maturation showed a significant variation in developmentally competent oocytes to that of incompetent ones. Along with this, the transcript was also found to have protein binding ability which was confirmed by RNA electrophoretic mobility shift assay. The protein used in the experiment was isolated from oocyte and cumulus cells via sonication. A novel lincRNA has been reported here that might have an important role in maturation of oocytes, inferred from its relative gene expression study and protein binding characteristics.
研究母源卵母细胞特异性基因是阐明调控卵母细胞质量、早期胚胎发育和存活的潜在机制的重要步骤。随着水牛卵母细胞特异性消减 cDNA 文库的建立，提出了一些需要回答的新问题。本研究从文库中选择了一个 EST 进行了特征描述，并强调了它在卵母细胞质量中的重要性。所选 EST 通过 RLM-RACE 进行全长扩增，鉴定出了 4 种转录变体。生物信息学分析表明全长转录本具有新颖性和保守的基因间特性。根据编码潜能计算器的输出，最大的转录本被鉴定为长基因间非编码 RNA。在卵母细胞成熟不同时间点的表达分析显示，具有发育能力的卵母细胞与不具有发育能力的卵母细胞之间存在显著差异。此外，还发现该转录本具有蛋白结合能力，这通过 RNA 电泳迁移率变动分析得到了证实。实验中使用的蛋白是通过超声处理从卵母细胞和卵丘细胞中分离得到的。根据相对基因表达研究和蛋白结合特性，本文报道了一种新的 lincRNA，它可能在卵母细胞成熟过程中发挥重要作用。
