Ordered gene knockout libraries for Neisseria gonorrhoeae

摘要 淋病奈瑟菌是性传播感染淋病的唯一病原体， 仅限于人类。N.淋病一直与人类宿主共同进化 所有现代历史，并已演变出与其发病模式相关的特定发病机制。 传播和它殖民的解剖部位。淋病发病率沿着快速上升 获得抗生素耐药性。因此，迫切需要了解N. 淋病发病机制，以开发新的治疗方案或候选疫苗。N.淋病 是天然的DNA转化能力，是少数几种细菌之一， 规范权限。这笔资源赠款将利用N的转型能力。 淋病和合成DNA构建体，以创建两个最重要的有序基因敲除文库， 常用的N。淋病菌株，MS 11和FA 1090。这些菌株代表了 细菌的系统发育分支。所述文库将包括编码一个或多个氨基酸的所有基因的缺失。 开放阅读框（ORF），其允许用非极性卡那霉素替换ORF 抗性基因（KmR）。每个KmR插入突变体还将携带独特的随机条形码。 该领域的主要研究人员和NIAID项目官员也支持这一资源提案， 这是一个重要的共享资源，有助于推动这一领域的进展。

Abstract Neisseria gonorrhoeae is the sole causative agent of the sexually transmitted infection gonorrhea that is restricted to the human population. N. gonorrhoeae has co-evolved with its human host throughout all modern history and has evolved specific mechanisms of pathogenesis that relate to its mode of transmission and the anatomical sites it colonizes. Rates of gonorrhea are rising along with a rapid acquisition of antibiotic resistance. There is an urgent need to understand of the mechanisms of N. gonorrhoeae pathogenesis to develop new treatment options or vaccine candidates. N. gonorrhoeae is naturally competent for DNA transformation and is one of a few bacterial species that does not regulate competence. This resource grant will leverage the transformation competence of N. gonorrhoeae and synthetic DNA constructs to create ordered gene knockout libraries of the two most commonly used N. gonorrhoeae strains, MS11 and FA1090. These strains represent different phylogenetic clades of the bacterium. The libraries will include deletions of all genes encoding an open reading frame (ORF) that will allow the replacement of the ORF with a non-polar kanamycin resistance gene (KmR). Each KmR insertion mutant will also carry a unique random barcode. Leading researchers in the field and NIAID program officials also support this resource proposal as an important shared resource to help advance progress in this field.