摘要
本研究为了探究广东省疥螨猪变种(Sarcoptes.scabiei var.suis)的基因型及与其他疥螨变种的进化关系,通过提取疥螨基因组DNA,利用PCR扩增、Sanger测序获取线粒体的ND1基因和核糖体DNA的ITS2基因,对两个目标基因核苷酸应用生物信息学分析工具BLAST和MEGA-X进行同源性分析并构建系统进化树。结果表明,所扩增广东猪疥螨的线粒体ND1基因序列大小为211 bp,1TS2 r DNA基因片段大小均为417 bp。在nt数据库疥螨亚目的所有序列中,本试验扩增的广东猪疥螨ND1基因序列与疥螨属ND1基因相似性最高(Identity>99%,E-value<6.20E-106),其次是户尘螨(Identity=80%,E-value=1.37E-44)、兔痒螨Psoroptes cuniculi(Identity=74%,E-value=9.84E-34)和粉尘螨Dermatophagoides farina(Identity=74%,E-value=4.18E-32)。本试验扩增的ITS2+序列与疥螨属ITS2基因序列相似性高(Identity为97.3%~99.3%,E-value=0),其次为背肛螨属(Identity为95.50%~97.80%,E-value为7.16E-35~4.83E-37)。通过序列比对与进化分析发现,在疥螨属内核糖体ITS2基因变异较小,线粒体ND1基因在疥螨属内有一定程度的变异。广东所采疥螨猪变种的ND1基因和ITS2基因与其他动物或人体内采集的疥螨相似程度高,疥螨ITS2基因在不同地域、不同宿主条件下的变异程度很小,在进行疥螨属内的遗传变异分析时不宜做目标分子,而通过对疥螨线粒体ND1基因分析发现其在疥螨属内存在一定的变异,可以作为疥螨属内遗传变异分析的候选分子。本研究可为今后疥螨的分类、鉴定及遗传变异分析提供一定依据。
To explore the genotype of Sarcoptes scabiei var.suis collected form in Guangdong Province and its evolutionary relationship with other S.scabiei from different animal and location.The mtDNA ND1 and r DNA ITS2 genes from swine were amplified through PCR technology using extracted genomic DNA as the template.The nucleotide sequence was analyzed by BLSAT program,and then use the MAGAX software for the homology analysis and phylogenetic tree construction.The results showed that the length of mtDNA ND1 and r DNA ITS2 gene was 211 bp and 417 bp,respectively.The partial mtDNA sequence has the highest similarity with some S.scabiei sequences(Identity>99%,E-value<6.20 E-106),followed by Dermatophagoides pteronyssinus(Identity=80%,E-value=1.37 E-44),Psoroptes cuniculi(Identity=74%,E-value=9.84 E-34)and Dermatophagoides farina(Identity=74%,E-value=4.18 E-32).In addition,the r DNA ITS2+sequence has the highest similarity with some S.scabiei sequences(Identity is 97.3%~99.3%,E-value=0),followed by the genus Notoedres(Identity is 95.50%~97.80%,E-value is 7.16 E-35~4.83 E-37),but has relative lower E-value in Sarcoptiforms.Last,the sequence similarity and molecular phylogenetic analysis showed that r DNA ITS2 gene has less variation and mtND1 gene has variation in the genus scabies.It is concluded that the ND1 and ITS2 genes has a high similarity with S.scabiei and the ITS2 gene is not suitable to be the target molecule when analyzing the genetic variation within the genus Scabies.Furthermore,this study gives a theoretical foundation for the further study on the molecular evolution of the genus scabies.
作者
黄福强
唐志强
李静
孙悦翔
钟秉洲
戚少贤
HUANG Fu-qiang;Tang Zhi-qiang;LI Jing;SUN Yue-xiang;ZHONG Bing-zhou;Qi Shao-xian(School of Life Science and Engineering,Foshan University,Foshan 528225,China)
出处
《特产研究》
2021年第5期19-25,共7页
Special Wild Economic Animal and Plant Research
基金
广东省自然科学基金(2018A1660004)
佛山科学技术学院高层次人才科研启动项目(CGZ07241)
广东省现代草牧业(羊)产业技术体系项目(2019KJ127)。
