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原肠胚形成期鹌鹑胚胎的电穿孔和增强型绿色荧光蛋白标记

Electroporation and EGFP labeling of gastrulating quail embryos.

作者信息

Cui Cheng, Lansford Rusty, Filla Mike B, Little Charles D, Cheuvront Tracey J, Rongish Brenda J

机构信息

Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, Kansas, USA.

出版信息

Dev Dyn. 2006 Oct;235(10):2802-10. doi: 10.1002/dvdy.20895.

Abstract

Labeling embryonic cells to trace their motion is a classical experimental approach with a host of techniques being used to mark live cells and tissues. Genetically engineered fluorescent protein vectors (DNA plasmids) are a recent technology well suited to time-resolved studies of cellular motion in live embryos. DNA plasmids encoding fluorescent proteins can be introduced into cells using several methods, including electroporation, a technique used widely for analysis of tissue culture and embryonic cells. Here we describe a technique designed to introduce DNA plasmids into early gastrulation stage quail embryos, ex ovo. The method is effective, and with practice enables an investigator to direct the vectors to relatively confined regions of gastrulating embryos. The required electroporation chamber can be fabricated from common laboratory materials. We anticipate that using this method of labeling cells in a warm-blooded embryo, during gastrulation, will be a fruitful means of studying subsequent embryogenesis.

摘要

标记胚胎细胞以追踪其运动是一种经典的实验方法,有许多技术可用于标记活细胞和组织。基因工程荧光蛋白载体(DNA质粒)是一项最新技术,非常适合对活胚胎中的细胞运动进行时间分辨研究。编码荧光蛋白的DNA质粒可通过多种方法导入细胞,包括电穿孔法,这是一种广泛用于组织培养和胚胎细胞分析的技术。在此,我们描述一种将DNA质粒导入原肠胚形成早期鹌鹑胚胎(体外)的技术。该方法有效,经过实践,研究人员能够将载体导入原肠胚胚胎相对局限的区域。所需的电穿孔室可由常见的实验室材料制成。我们预计,在原肠胚形成期间,使用这种方法标记温血胚胎中的细胞将是研究后续胚胎发育的一种富有成效的手段。

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