收稿日期: 2022-01-24
修回日期: 2022-04-09
网络出版日期: 2022-08-02
基金资助
国家自然科学基金(81760607);国家自然科学基金(U2002219);云南省自然科学基金(2017FD139);云南省教育厅科学研究基金(2022J0687)
Complete mitochondrial genome sequence of Rhipicephalus microplus
Received date: 2022-01-24
Revised date: 2022-04-09
Online published: 2022-08-02
Supported by
National Natural Science Foundation of China(81760607);National Natural Science Foundation of China(U2002219);Yunnan Natural Science Foundation(2017FD139);Scientific Research Fund of Yunnan Education Department(2022J0687)
2017年9月采用布旗法在云南省临沧市耿马县采集游离的微小扇头蜱,提取蜱DNA,采用通用引物PCR扩增微小扇头蜱的16S rRNA、12S rRNA、nad1、cox3基因片段,采用长PCR扩增上述4个基因区的长片段,产物纯化后采用步移法测序,拼接获得线粒体基因组全序列。结果显示,线粒体基因组全长14 763 bp,包含13个蛋白质编码基因、2个rRNA基因、22个tRNA和1个长度为264 bp的非编码区（D-Loop区）,碱基组成为A 38.66%、G 9.02%、C 11.46%、T 40.87%,（A + T）79.53%。蛋白质编码基因起始密码子均为ATN;除COX2、cox3、NAD5使用不完整T作为终止密码子外,其余基因终止密码子均为TAA或TAG;基因重叠区有12处共95 bp,长度在1~24 bp;基因间隔区有18处共433 bp,长度在1~311 bp。22个tRNA中,除tRNACys、tRNAPhe、tRNASer以外均能形成典型的三叶草结构。
刘雅芳 , 陈彬 , 芦新焱 , 李光华 , 杜春红 , 姜丹丹 , 杨兴 . 云南微小扇头蜱线粒体基因组全序列测定与分析[J]. 中国寄生虫学与寄生虫病杂志, 2022 , 40(5) : 677 -681 . DOI: 10.12140/j.issn.1000-7423.2022.05.018
Rhipicephalus microplus was collected by the cloth flag method in Yunnan Province in September 2017 and was used for DNA extraction. Four long fragments were amplified by long PCR using the primers designed based on 16S rRNA, 12S rRNA, nad1 and cox3 gene sequences. The fragments were sequenced by conserved primer-walking. The complete mitochondrial genome sequence was obtained by splicing. The length of the mitochondrial genome (mtDNA) of Rhipicephalus microplus is 14 763 bp. It contained 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and a non-coding control region of 264 bp (D-Loop region). The base composition was A 38.66%, G 9.02%, C 11.46%, T 40.87%. A + T content was 79.53%. The starting codon of protein-coding genes was ATN. The stop codons were TAA or TAG, except for cox2, cox3, nad5, which used incomplete T as the stop codon. The complete mitochondrial genome have 12 gene overlaps. The length ranges from 1-24 bp. The length of 18 total intergenic regions was 433 bp ranging from 1-311 bp. A total of 22 transferring RNA were found, all of which were typical cloverleaf structures except for tRNACys, tRNAPhe, tRNASer.
Key words： Rhipicephalus microplus; Mitochondrial genome; Sequence analysis
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