Biology Department, The American University in Cairo , New Cairo , Egypt.
Biology Department, The American University in Cairo , New Cairo , Egypt ; Biotechnology Program, The American University in Cairo , New Cairo , Egypt.
PeerJ. 2014 Dec 4;2:e689. doi: 10.7717/peerj.689. eCollection 2014.
A number of polymerase chain reaction (PCR) inhibitors have been identified from biological and environmental samples. By and large, such substances are treated as random nuisances and contaminants with alternate functions; their inhibitory effects on DNA replication being a coincidental property of their molecular structure. Here, we demonstrate the presence of a localized PCR inhibitor in the foregut of the porcelain crab Petrolisthes rufescens (Anomura: Porcellanidae) from the Red Sea. The inhibitor precluded amplification of 28s, 16s and 18s gene sequences effectively but lost activity at 10(-2) dilutions from initial concentration. Heat treatment was ineffective in arresting inhibition and spectrophotometric techniques suggested that the inhibitor was not a melanin-type compound. The compound was not detected from midgut, hindgut, or gills of the crab. Activity of the inhibitor was precluded when samples were treated with suspensions from the midgut, suggesting that enzymatic degradation of the inhibitor likely happens at that part of the gut. As many microbial pathogens invade their hosts via ingestion, we suggest the presence of the localized inhibitor could carry a defensive or immunological role for P. rufescens. The identity of the inhibitory molecule remains unknown.
已从生物和环境样本中鉴定出多种聚合酶链反应 (PCR) 抑制剂。总的来说,这些物质被视为随机的滋扰物和具有替代功能的污染物;其对 DNA 复制的抑制作用是其分子结构的偶然特性。在这里,我们证明了来自红海的瓷蟹 Petrolisthes rufescens(Anomura:Porcellanidae)前肠中存在局部 PCR 抑制剂。该抑制剂有效地阻止了 28s、16s 和 18s 基因序列的扩增,但在初始浓度的 10(-2) 稀释度下失去活性。热处理不能阻止抑制作用,分光光度技术表明抑制剂不是黑色素型化合物。该化合物未从蟹的中肠、后肠或鳃中检测到。当用中肠悬浮液处理样品时,抑制剂的活性被阻止,这表明抑制剂可能在肠道的那个部位被酶降解。由于许多微生物病原体通过摄入入侵宿主,我们认为局部抑制剂的存在可能对 P. rufescens 具有防御或免疫作用。抑制分子的身份仍然未知。