嗜热四膜虫表达载体的优化

Optimization of Tetrahymena thermophila Expression Vector

近年来淡水纤毛虫嗜热四膜虫成为新的重组亚单位疫苗生产平台。为了进一步提高重组四膜虫的筛选效率,使其能更广泛地应用于重组亚单位疫苗制备,本研究利用基因工程手段,改造了原有的表达载体,把带有巴龙霉素抗性的neo基因换成嘌呤霉素抗性表达盒,从而提高重组四膜虫的筛选效率,达到优化表达载体的目的,同时将常用的氯化镉诱导的MTT1启动子替换成HSP70启动子,实现热诱导表达,为嗜热四膜虫表达平台在重组亚单位疫苗的进一步应用奠定基础。

The freshwater ciliate, Tetrahymena thermophila, has recently emerged as a novel manufacturing platform for recombinant subunit vaccines. We aim to further improve the screening efficiency of recombinant Tetrahymena to make it more widely used in the preparation of recombinant subunit vaccine. In this study, we constructed a new vector with the codon optimized puromycin resistance cassette replaced paromomycin resistance cassette to improve the screen efficiency and make the vector more optimum for expression. We also substitute thermo-inducible promoter HSP70 for MTT1 promoter which was induced by CdCI~. The expression of foreign proteins was triggered simply by increasing the cultural temperature. Construction of this vector would widen the application of T. thermophila expression system to preparation of more suhunit vaccines.