Gene expression in commensal tsetse symbionts

描述（由申请人提供）：人类非洲昏睡病是一种由锥虫（布鲁氏锥虫）引起的潜在致命疾病。抑制这种迅速再次出现的疾病传播的核心是控制节肢动物媒介采采蝇（舌蝇属）。采采含有一种共生的内共生微生物（Sodalis舌虫），生活在靠近锥虫的中肠中。本研究的目的是利用这种共生菌群来降低采采蝇传播锥虫的能力。为了实现这一目标，我计划改进现有的基于染色体外质粒的转化程序，通过同源重组将“效应分子”基因整合到Sodalis的染色体中。有两种候选分子，具有证明的锥虫降解活性，可用于此目的：采采附合素和脊椎动物BMAP-27。重组Sodalis将重新引入采采蝇。然后将监测这些“准转基因”果蝇及其后代在适应性、重组蛋白表达的稳定性和锥虫病载体能力方面的变化。

DESCRIPTION (provided by applicant): African sleeping sickness in humans is a potentially deadly disease caused by trypanosomes (Trypanosoma brucei spp.). Central to inhibiting the spread of this rapidly re-emerging disease is control of the arthropod vector, the tsetse fly (genus Glossina). Tsetse harbors a commensal endosymbiotic microorganism (Sodalis glossinidius) that lives in the midgut in close proximity to the trypanosomes. The goal of this research is to exploit this symbiotic flora for the purpose of decreasing tsetse's capacity to transmit trypanosomes. To accomplish this goal, I plan to improve upon an existing extra-chromosomal plasmid-based transformation procedure by using homologous recombination to incorporate 'effector molecule' genes into Sodalis' chromosome. 2 candidate molecules, with demonstrated trypanolytic activity, are available for this purpose: tsetse attacin and vertebrate BMAP-27. Recombinant Sodalis will be reintroduced to tsetse. These 'paratransgenic' flies and their offspring will then be monitored for changes in fitness, stability of recombinant protein expression and trypanosome vectorial capacity.