请阅读以下文献,思考后回答下列问题: Moreira LA, et al.A Wolbachia Symbiont in Aedes aegypti Limits Infection with Dengue,Chikungunya, and Plasmodium Cell 2009;139(7):1268–78 1) 在 1269 页左半页最下面一段 , These mosquito strains were compared to tetracycline-treatedcounterparts that were genetically identical but lacked the Wolbachiainfection, named PGYP1.tet and PGYP1.out.tet, respectively. 用四环素处理得到的蚊作为 Wolbachia 阴性样本有何潜在问题?实验设计上可以做何改进? 2) 在 1269 页右半页第二段 , To provide a more conservative test ofWolbachia-mediated interference, mosquitoes were intrathoracically injectedwith DENV-2. These experiments circumvented the midgut barrier to infection(Woodring et al., 1996) and allowed for the delivery of a repeatableinoculating dose (around 2,750 infectious particles/mosquito) of DENV-2 thatproduced consistent high-titer infections in control mosquitoes. 这种胸内直接注射的方法有何优点?有何潜在的问题?是否人为地将可能影响 Dengue 感染的因素排除在考虑之外?这些被排除考虑的因素可能有哪些? 3) 检测全身的总病毒量后,为何还要分部位检测病毒量?哪个检测指标跟疾病的传播更有相关性? 4) 文章中检测了免疫相关的基因表达,并提出 Wolbachia 可能通过激活免疫应答而影响 Dengue, Chikungunya 以及 Plasmodium 的传播。你认为这一假说是否合理? Wolbachia 作为稳定存在于蚊体内的共生菌,是否可能被免疫系统识别?作为对比,转基因小鼠通常会对转基因表达的抗原产生免疫应答吗?