Movila Alexandru, Morozov Alexandr, Sitnicova Natalia
Institute of Zoology, Moldova Academy of Sciences, 1 Academiei str., Chisinau, Republic of Moldova.
J Parasitol. 2013 Feb;99(1):40-3. doi: 10.1645/GE-3225.1. Epub 2012 Aug 27.
The molecular genetic variability of the 12S rRNA gene, on the basis of partial primary sequence and in silico-predicted secondary structures among Dermacentor reticulatus (Fabricius 1794) ticks, was studied in the Chernobyl Nuclear Power Plant (ChNPP) Exclusion Zone. In total, 20, 20, and 25 ethanol-preserved specimens, previously collected at 3 sites with 0.76, 1.91, and 4.5 millisievert (mSv)/hr ionizing radiation background, were examined. The primary sequence analysis generated 4 haplotypes defined by 3 polymorphic sites. The most common haplotype 1 was found in all 3 locations, representing 86.2% of all sampled individuals. Haplotype 4 (10.8%) was detected at the 1.91 and 4.50 mSv/hr sites. The unique haplotypes 2 (1.5%) and 3 (1.5%) were detected only at the 1.91 and 4.50 mSv/hr sites, respectively. The haplotype diversity, nucleotide diversity, and pairwise nucleotide differences for 2 tick populations at the 1.90 and 4.50 mSv/hr sites were 0.279, 0.00085, and 0.289, and 0.397, 0.00122, 0.413, respectively. No polymorphism was detected in ticks collected at the 0.76 mSv/hr site. The primary sequences of 12S rRNA were folded into the secondary structures and the free energy of haplotypes was calculated. The free energy at 37 C (ΔG) of the nonmutant haplotype 1 and the mutant haplotypes 2, 3, and 4 were -45.79, -44.17, -39.56, and -45.79 kcal/mol, respectively. Considering the correlation between the structural profile similarity of 12S rRNA and point mutations, haplotypes 1 and 4 have similar secondary structure profiles and have received a 0.999219 similarity score in the cluster tree. The unique haplotypes 2 and 3 have differences in the secondary structure in comparison with haplotypes 1 and 4; the similarity scores were 0.914747 and 0.169431, respectively. Further studies using more genetic markers are warranted to ascertain the genetic variability and population genetic structure within D. reticulatus tick populations in the ChNPP Exclusion Zone and to resolve their vector capacity.
在切尔诺贝利核电站(ChNPP)禁区内,基于部分一级序列以及网纹革蜱(Dermacentor reticulatus,Fabricius 1794)蜱虫的计算机预测二级结构,对12S rRNA基因的分子遗传变异性进行了研究。总共检查了20只、20只和25只保存在乙醇中的标本,这些标本之前分别采集于3个地点,其电离辐射背景分别为0.76、1.91和4.5毫希沃特(mSv)/小时。一级序列分析产生了由3个多态性位点定义的4种单倍型。最常见的单倍型1在所有3个地点均有发现,占所有采样个体的86.2%。在1.91和4.50 mSv/小时的地点检测到单倍型4(10.8%)。独特的单倍型2(1.5%)和3(1.5%)分别仅在1.91和4.50 mSv/小时的地点检测到。1.90和4.50 mSv/小时地点的2个蜱虫种群的单倍型多样性、核苷酸多样性和成对核苷酸差异分别为0.279、0.00085和0.289,以及0.397、0.00122和0.413。在0.76 mSv/小时地点采集的蜱虫中未检测到多态性。将12S rRNA的一级序列折叠成二级结构并计算单倍型的自由能。非突变单倍型1以及突变单倍型2、3和4在37℃时的自由能(ΔG)分别为-45.79、-44.17、-39.56和-45.79千卡/摩尔。考虑到12S rRNA的结构轮廓相似性与点突变之间的相关性,单倍型1和4具有相似的二级结构轮廓,并且在聚类树中获得了0.999219的相似性评分。与单倍型1和4相比,独特的单倍型2和3在二级结构上存在差异;相似性评分分别为0.914747和0.169431。有必要使用更多的遗传标记进行进一步研究,以确定ChNPP禁区内网纹革蜱种群内的遗传变异性和群体遗传结构,并解析它们的传播能力。