Yasuoka Yuuri, Taira Masanori
Marine Genomics Unit, Okinawa Institute of Science and Technology Graduate University, Onna-son, Okinawa 904-0495, Japan.
Department of Biological Sciences, Graduate School of Science, University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan
Cold Spring Harb Protoc. 2019 Jan 2;2019(1):pdb.prot097279. doi: 10.1101/pdb.prot097279.
Introducing exogenous DNA into an embryo can promote misexpression of a gene of interest via transcription regulated by an attached enhancer-promoter. This protocol describes plasmid DNA microinjection into embryos for misexpression of genes after zygotic gene expression begins. It also describes a method for coinjecting a reporter plasmid with mRNA or antisense morpholinos to perform luciferase reporter assays, which are useful for quantitative analysis of -regulatory sequences responding to endogenous or exogenous stimuli in embryos.
将外源DNA导入胚胎可通过附着的增强子-启动子调控的转录促进目的基因的错误表达。本方案描述了在合子基因表达开始后，将质粒DNA显微注射到胚胎中以实现基因的错误表达。它还描述了一种将报告质粒与mRNA或反义吗啉代寡核苷酸共注射以进行荧光素酶报告基因检测的方法，这对于定量分析胚胎中对内源或外源刺激作出反应的调控序列很有用。
