一种用于生物膜中基因水平转移研究的微流控方法

A microfluidic method applied to observe gene transfer in biofilm

职称论文发表摘要： 目前，用于研究基因水平转移的实验方法主要有基于培养的方法、基于分子生物学的方法和基于荧光的方法，但这些方法都很难实现对于基因转移过程的实时和原位观测。本研究设计了一种新的实验方法，利用接种有供体菌和受体菌的微流控芯片作为实验载体，并通过引入基因改造的供体菌，实现实时原位观察基因水平转移的过程，并可对转移频率和转移结果加以分析。通过对基因转移图像处理进行计算得到转移率为0.18%-2.37%，流式细胞分析结果表明转化结合子的比例为0.2%。

关键词： 微流控芯片；基因水平转移；实时观测

ZHANG Jing1,， LI Bing1,， QIU Yong2,*

（  1、School of Environment,Tsinghua University,Beijing 100084；       2、School of Energy and environmental engineering,University of Science and Technology, Beijing 100084；  ）

Abstract： ICurrently, methods applied to the researches of gene transfer are mainly based on culture, molecular biology or fluorescence. However, the methods above are not capable to realize real-time observation and to record gene transfer process.In this study, we design a novel method that inoculating donor and receptor on a single-channel microfluidic chip and the donor strain induced was geneticallly modified. The method can realize in-situ and real-time observation of the process of gene transfer, and the transfer rate and results can be analysed. The transfer rate calculated from processed images is 0.18%-2.37%, and the proportion of transconjugants got through flow cytometry analysis is 0.2%.

Keywords： microfluidic chip;honrizontal gene transfer;real-time observation