一种新型的Tet-off慢病毒调控系统的构建及其调控作用

A Novel Tetracycline-Suppressed Expression System Based on Single Lentiviral Vector

四环素调控的真核表达系统是真核细胞诱导表达系统的重要组成部分。当前大多数研究者采用双质粒四环素表达调控系统调控外源基因的表达。由于,双质粒四环素表达调控系统的基因表达调控严格依赖于两个单载体同时感染同一个细胞,因而限制其在体内实验中的效率。为解决该问题,研究构建了由慢病毒载体构成的单质粒四环素表达调控系统-pLV300(Tet-off)。构建的载体经酶切鉴定及测序均正确。通过荧光观察及RT-PCR实验结果显示,该单质粒四环素调控的慢病毒载体体外可以高效地转移并且维持目的基因的表达,其表达调控效应高、无明显毒副作用,且调控严密。该表达调控系统为慢病毒的临床应用奠定了一定基础。

Tetracycline-regulated eukaryotic expression system is a useful tool in area of biological researches.However,the current system is characterized of separation of essential elements into two individual vectors,resulting in inconvenience in the process of application.Here,the authors successfully generate a single-vector tetracycline-regulated expression system which is based on recombinant lentivirus.Our data confirmes the efficiency and reliability of this single-vector controllable expression system.It is hoped that the work would advance the application of tetracycline-regulated system in future.