中文摘要：转基因动物的分子特征对于其性状分析、非预期效果预测、商业需求和经济需求来说至关重要。本文用利用新一代测序（NGS）方法对hCD46转基因猪的综合分子特征进行了研究。由于NGS成本高于其他分子方法，因而为了降低成本且使测序策略更经济有效，作者采用计算机模拟识别能满足分子特征分析需求的最小数据集。本文获得的分子特征信息为转基因动物的安全性评价奠定了坚实的基础。研究结果表明，新一代测序（NGS）方法可用于转基因生物分子特征分析和安全评价的可靠准确方法。
外文摘要：Molecular characterization of transgenic animals is crucial for the phenotype analysis, unintended effects prediction, commercial and economic requirements. In this study, a comprehensive molecular characterization of a transgenic pig, which expressed hCD46 gene, was verified using next-generation sequencing (NGS). One complete and one incomplete sequence of pBSCD46-neo and multiple backbone fragments were inserted into the host genome. The whole insertion sequence was 28,243 bp, and 12 bp sequence on the genome was deleted during the transgene. The cost of NGS is higher than other molecular methods, in order to reduce the cost and to make the sequencing strategy more economically effective, we carried out a computer simulation to identify the smallest amount of data that could meet the requirements of the molecular characterization analysis. 15 x depth of coverage was shown to be sufficient with good accuracy, while 3 x might be effective, 20 x was recommended. The obtained molecular characterization information would provide a stronger foundation for the safety evaluation of the GM animals. Meanwhile, the gene integrity of the porcine pathogen PERV was analyzed using the same procedure. No complete sequence of the PERV subtype was observed in the porcine genome, which was the same as the previous research. In consequence, NGS combined bioinformatics analysis is a reliable and accurate approach for the molecular characterization of GMOs, even for the safety evaluation.
外文关键词：Transgenic pig; hCD46; Molecular characterization; Next-generation sequencing; Sequencing depth; PERV
作者：Wang, Qin; Shi, Ningning; Shang, Ying; 等.
作者单位：Chinese Acad Inspect & Quarantine
期刊名称：GENE
期刊影响因子：2.138
出版年份：2017
出版刊次：8
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