Programa de Pós-Graduação em Biotecnologia e Recursos Naturais (PPG-MBT), Universidade Estadual do Amazonas - UEA, Manaus, AM, Brazil.
Coordenação de Biodiversidade (COBIO), Laboratório Temático de Biologia Molecular (LTBM), Programa de Pós-Graduação em Genética, Conservação e Biologia Evolutiva (PPG-GCBEv), Instituto Nacional de Pesquisas da Amazônia (INPA), Manaus, AM, Brazil.
Mol Biol Rep. 2023 Aug;50(8):7099-7104. doi: 10.1007/s11033-023-08567-2. Epub 2023 Jun 14.
Anopheles darlingi is a monotypic species in terms of its morphological, genetic, and behavioral aspects and is the primary transmitter of human malaria (99%) in Brazil, especially in the Brazilian Amazon. In this pioneering study, 15 expressed sequence tag (EST)-simple sequence repeat (SSR) markers were obtained and characterized in samples from the municipality of São Gabriel da Cachoeira, Amazonas state, Brazil, with polymorphisms that can be used for further genetic research.
The specimens (from egg to larval stage) collected were bred in the insectary at INPA (National Institute for Amazonian Research). The SSR repeats within the contigs of the A. darlingi EST banks were confirmed on the Vector Base site. DNA was extracted and amplified using polymerase chain reaction and then genotyped. Fifteen polymorphic SSR loci were identified and characterized. The number of alleles totaled 76 and ranged from 2 to 9. The observed heterozygosity varied between 0.026 and 0.769, the expected heterozygosity between 0.025 and 0.776, and the mean polymorphism information content was 0.468. Eight loci showed Hardy-Weinberg equilibrium (HWE) after Bonferroni correction (P: (5%) ≤ 0.0033). No linkage disequilibrium was found among the loci.
The polymorphic SSRs of the loci have been shown to be efficient for investigation of the variability and genetic population structure of A. darlingi.
在形态、遗传和行为方面，冈比亚按蚊是一个单型种，是巴西（尤其是在巴西亚马孙地区）人类疟疾（99%）的主要传播媒介。在这项开创性的研究中，从巴西亚马逊州圣加布里埃尔达卡恰乌埃拉市的样本中获得并鉴定了 15 个表达序列标签（EST）-简单重复序列（SSR）标记，这些标记具有多态性，可用于进一步的遗传研究。
在 INPA（国家亚马孙研究所）的昆虫学实验室中饲养收集的（从卵到幼虫阶段）标本。在 Vector Base 网站上确认了 A. darlingi EST 库的序列内 SSR 重复。使用聚合酶链反应提取和扩增 DNA，然后进行基因分型。鉴定并描述了 15 个多态性 SSR 位点。等位基因总数为 76 个，范围为 2 到 9。观察到的杂合度在 0.026 到 0.769 之间变化，预期杂合度在 0.025 到 0.776 之间变化，平均多态信息含量为 0.468。经过 Bonferroni 校正（P：（5%）≤0.0033），8 个位点显示 Hardy-Weinberg 平衡（HWE）。未发现这些位点之间的连锁不平衡。
这些多态性 SSR 位点对于冈比亚按蚊的变异性和遗传种群结构的研究是有效的。
