Comprehensive molecular characterization of a transgenic pig expressing hCD46 gene

文献类型： 外文期刊

第一作者： Wang, Qin

作者： Wang, Qin;Liu, Xiaofei;Fu, Wei;Lin, Xiangmei;Shi, Ningning;Pan, Dengke;Shi, Ningning;Shang, Ying;Shang, Ying;Xu, Wentao;Zhao, Yiqiang

作者机构：

关键词： Transgenic pig;hCD46;Molecular characterization;Next-generation sequencing;Sequencing depth;PERV

期刊名称：GENE （ 影响因子：3.688； 五年影响因子：3.329 ）

ISSN：

年卷期：

页码：

收录情况： SCI

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.

分类号： R394

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