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海胆胚胎中调控基因的功能研究。

Functional studies of regulatory genes in the sea urchin embryo.

作者信息

Cavalieri Vincenzo, Di Bernardo Maria, Spinelli Giovanni

机构信息

Dipartimento di Biologia Cellulare e dello Sviluppo "A. Monroy", Università di Palermo, Palermo, Italy.

出版信息

Methods Mol Biol. 2009;518:175-88. doi: 10.1007/978-1-59745-202-1_13.

Abstract

Sea urchin embryos are characterized by an extremely simple mode of development, rapid cleavage, high transparency, and well-defined cell lineage. Although they are not suitable for genetic studies, other approaches are successfully used to unravel mechanisms and molecules involved in cell fate specification and morphogenesis. Microinjection is the elective method to study gene function in sea urchin embryos. It is used to deliver precise amounts of DNA, RNA, oligonucleotides, peptides, or antibodies into the eggs or even into blastomeres. Here we describe microinjection as it is currently applied in our laboratory and show how it has been used in gene perturbation analyses and dissection of cis-regulatory DNA elements.

摘要

海胆胚胎的特点是发育模式极其简单、卵裂迅速、透明度高且细胞谱系明确。尽管它们不适合进行遗传学研究,但其他方法已成功用于揭示参与细胞命运特化和形态发生的机制及分子。显微注射是研究海胆胚胎基因功能的首选方法。它用于将精确数量的DNA、RNA、寡核苷酸、肽或抗体导入卵甚至卵裂球中。在此,我们描述目前在我们实验室中应用的显微注射方法,并展示它如何用于基因干扰分析和顺式调控DNA元件的剖析。

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