Xu Xing-Li, Yang Hu
College of Life Sciences and Resource Environment, Yichun University, Yichun, Jiangxi Province, China.
J Med Entomol. 2021 Nov 9;58(6):2540-2546. doi: 10.1093/jme/tjab139.
Rhipicephalus microplus is the main blooding-sucking ectoparasite of bovines and is regarded as important vectors of animal diseases such as Babesiosis. Mining protective antigens of R. microplus to develop antitick vaccine is the most potential tick control strategy. In this study, the specific primers were designed according to the conserved nucleotide sequence of enolase gene in Haemaphysalis flava, Ixodes ricinus, and Ornithodoros moubata. The fragment of enolase gene was obtained by PCR using cDNA template from fully engorged female R. microplus. The full-length cDNA of enolase gene was amplified using rapid amplification of cDNA ends (RACE). Expression pattern of enolase gene in different tissues of R. microplus was analyzed by real-time quantitative PCR (qRT-PCR). Results showed that the full-length enolase cDNA containing 2052 bp was obtained successfully. The complete cDNA included an ORF of 1305 nucleotides encoding a protein of 434 amino acids. The enolase exhibited 85.0% amino acid identity to the enolase of H. flava, 81.1% to I. ricinus enolase, and 81.3% to O. moubata enolase. qRT-PCR analysis indicated that the enolase had the highest expression in the salivary gland of R. microplus.
微小扇头蜱是牛的主要吸血外寄生虫，被视为巴贝斯虫病等动物疾病的重要传播媒介。挖掘微小扇头蜱的保护性抗原以开发抗蜱疫苗是最具潜力的蜱虫控制策略。在本研究中，根据黄褐血蜱、蓖麻硬蜱和波斯锐缘蜱烯醇化酶基因的保守核苷酸序列设计了特异性引物。以饱血雌蜱微小扇头蜱的cDNA为模板，通过PCR获得烯醇化酶基因片段。利用cDNA末端快速扩增（RACE）技术扩增烯醇化酶基因的全长cDNA。通过实时定量PCR（qRT-PCR）分析烯醇化酶基因在微小扇头蜱不同组织中的表达模式。结果表明，成功获得了长度为2052 bp的烯醇化酶全长cDNA。完整的cDNA包含一个1305个核苷酸的开放阅读框，编码一个434个氨基酸的蛋白质。该烯醇化酶与黄褐血蜱烯醇化酶的氨基酸同一性为85.0%，与蓖麻硬蜱烯醇化酶的同一性为81.1%，与波斯锐缘蜱烯醇化酶的同一性为81.3%。qRT-PCR分析表明，烯醇化酶在微小扇头蜱唾液腺中的表达最高。
