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利用目标捕获技术对小泰勒虫进行全基因组测序。

Whole genome sequencing of Theileria parva using target capture.

机构信息

Agricultural Research Council, Onderstepoort Veterinary Research, Private Bag X05, Onderstepoort, 0110 Pretoria, South Africa; Department of Veterinary Tropical Diseases, Vector and Vector-borne Disease Research Programme, University of Pretoria, Private Bag X04, Onderstepoort, 0110 Pretoria, South Africa.

Agricultural Research Council, Biotechnology Platform, Private Bag X05, Onderstepoort, 0110 Pretoria, South Africa.

出版信息

Genomics. 2021 Mar;113(2):429-438. doi: 10.1016/j.ygeno.2020.12.033. Epub 2020 Dec 25.

Abstract

Protozoan parasite isolation and purification are laborious and time-consuming processes required for high quality genomic DNA used in whole genome sequencing. The objective of this study was to capture whole Theileria parva genomes directly from cell cultures and blood samples using RNA baits. Cell culture material was bait captured or sequenced directly, while blood samples were all captured. Baits had variable success in capturing T. parva genomes from blood samples but were successful in cell cultures. Genome mapping uncovered extensive host contamination in blood samples compared to cell cultures. Captured cell cultures had over 81 fold coverage for the reference genome compared to 0-33 fold for blood samples. Results indicate that baits are specific to T. parva, are a good alternative to conventional methods and thus ideal for genomic studies. This study also reports the first whole genome sequencing of South African T. parva.

摘要

原生动物寄生虫的分离和纯化是一个繁琐且耗时的过程,需要使用高质量的基因组 DNA 进行全基因组测序。本研究的目的是使用 RNA 诱饵直接从细胞培养物和血液样本中捕获完整的小泰勒虫基因组。细胞培养物材料直接进行诱饵捕获或测序,而血液样本则全部进行捕获。诱饵在从血液样本中捕获小泰勒虫基因组方面的成功率各不相同,但在细胞培养物中是成功的。与细胞培养物相比,基因组图谱显示血液样本中宿主污染程度更高。与血液样本的 0-33 倍相比,捕获的细胞培养物对参考基因组的覆盖率超过 81 倍。结果表明,诱饵是小泰勒虫特有的,是传统方法的良好替代品,因此非常适合基因组研究。本研究还报告了南非小泰勒虫的首次全基因组测序。

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