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Sequence analysis of the cytochrome c oxidase subunit 1 gene of Sarcoptes scabiei isolated from goats and rabbits in East Java, Indonesia

机译:印度尼西亚东爪哇省山羊和兔分离的Sarcoptes scabiei的细胞色素c氧化酶亚基1基因的序列分析

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Aim: This study aimed to sequence the Cytochrome c oxidase (COX-1) gene sequence from mitochondrial DNA of Sarcoptes scabiei isolated from Lamongan goats and Mojokerto rabbits, align it with DNA isolated from Zi'gong rabbit (GenBank accession No. EU256389.1), and produce a phylogenetic analysis of S. scabiei COX-1 gene. Materials and Methods: S. scabiei mites were obtained from goats and rabbits, and DNA was extracted using QIAamp DNA Mini Kit. The forward and reverse primer sequences were designed based on the DNA sequence of an S. scabiei COX-1 gene isolated from the Zi'gong rabbit (5'-TCTTAGGGGCTGGATTTAGTATG-3' and 5'-AGTTCCTCTACCAGTTCCAC-3', respectively). To confirm sequencing output, the sequence resulting from the reverse primer was inverted and aligned to the sequence from the forward primer using Clone Manager Professional Version 9 for Windows (Scientific & Educational Software; http://www.scied.com). This alignment was subsequently used to build a phylogenetic tree, using the Neighbor- Joining method, in the MEGA6 program (https://www.megasoftware.net/). Results: Polymerase chain reaction (PCR) products from S. scabiei isolates from Lamongan goats and Mojokerto rabbits produced bands of around 290 bp with 2% agarose gel electrophoresis. Comparing the DNA sequences of the S. scabiei COX-1 gene with those isolated from Lamongan goats and Mojokerto rabbits showed 99% homology. Conclusion: PCR products of the S. scabiei COX-1 gene isolated from Lamongan goats and Mojokerto rabbits were around 290 bp long. The sequences had more than 99% homology. The sequences of the COX-1 gene of S. scabiei from Lamongan goats and Mojokerto rabbits were relatively close to the sequence of the gene in S. scabiei obtained from various hosts according to National Center for Biotechnology Information data.
机译:目的:本研究旨在对从Lamongan山羊和Mojokerto兔分离的Sarcoptes scabiei线粒体DNA的细胞色素c氧化酶(COX-1)基因序列进行测序,并将其与从自贡兔分离的DNA进行比对(GenBank登录号EU256389.1) ),并进行S. scabiei COX-1基因的系统发育分析。材料和方法:从山羊和兔子身上获得sc链霉菌螨,并使用QIAamp DNA Mini Kit提取DNA。正向和反向引物序列是基于从子公兔中分离的S. scabiei COX-1基因的DNA序列设计的(分别为5'-TCTTAGGGGCTGGATTTAGTATG-3'和5'-AGTTCCTCTACCAGTTCCAC-3')。为了确认测序输出,使用适用于Windows的Clone Manager专业版9(Scientific&Educational Software; http://www.scied.com),将反向引物产生的序列反转并与正向引物序列进行比对。该比对随后用于在MEGA6程序(https://www.megasoftware.net/)中使用Neighbor-Joining方法建立系统发育树。结果:来自Lamongan山羊和Mojokerto兔的sc。scabiei分离株的聚合酶链反应(PCR)产品通过2%琼脂糖凝胶电泳产生了约290 bp的条带。比较S. scabiei COX-1基因的DNA序列与从Lamongan山羊和Mojokerto兔分离的DNA序列,发现有99%的同源性。结论:从Lamongan山羊和Mojokerto兔分离的S. scabiei COX-1基因的PCR产物长约290 bp。序列具有超过99%的同源性。根据国家生物技术信息中心的数据,来自拉蒙根山羊和Mojokerto兔的斯卡宾链球菌COX-1基因的序列与从各种宿主获得的斯卡宾链球菌的基因序列相对接近。

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