Department of Medical Entomology, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand.
Dev Comp Immunol. 2012 Jan;36(1):104-11. doi: 10.1016/j.dci.2011.06.010. Epub 2011 Jun 28.
A cDNA encoding a lysozyme was obtained by rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR) from females of the malaria vector Anopheles dirus A (Diptera: Culicidae). The 623 bp lysozyme (AdLys c-1) cDNA encodes the 120 amino acid mature protein with a predicted molecular mass of 13.4 kDa and theoretical pI of 8.45. Six cysteine residues and a potential calcium binding motif that are present in AdLys c-1 are highly conserved relative to those of c-type lysozymes found in other insects. RT-PCR analysis of the AdLys c-1 transcript revealed its presence at high levels in the salivary glands both in larval and adult stages and in the larval caecum. dsRNA mediated gene knockdown experiments were conducted to examine the potential role of this lysozyme during Plasmodium berghei infection. Silencing of AdLys c-1 resulted in a significant reduction in the number of oocysts as compared to control dsGFP injected mosquitoes.
通过快速扩增 cDNA 末端聚合酶链反应 (RACE-PCR),从疟蚊 Anopheles dirus A 的雌性个体中获得了编码溶菌酶的 cDNA。该 623bp 的溶菌酶 (AdLys c-1) cDNA 编码 120 个氨基酸的成熟蛋白,预测分子量为 13.4kDa,理论等电点为 8.45。与其他昆虫中发现的 c 型溶菌酶相比,AdLys c-1 中的 6 个半胱氨酸残基和一个潜在的钙结合基序高度保守。AdLys c-1 转录本的 RT-PCR 分析表明,在幼虫和成虫的唾液腺以及幼虫的盲肠中均存在高水平的表达。dsRNA 介导的基因敲低实验用于研究该溶菌酶在伯氏疟原虫感染期间的潜在作用。与对照 dsGFP 注射的蚊子相比,AdLys c-1 的沉默导致卵囊数量显著减少。