Department of Biology, New York City College of Technology of the City University of New York, Brooklyn, NY, USA.
Department of Biology, Brooklyn College of The City University of New York, Brooklyn, NY, USA.
Methods Mol Biol. 2022;2364:349-361. doi: 10.1007/978-1-0716-1661-1_18.
Biolistic bombardment is widely used as a means of delivering vector-coated microparticles into microorganisms, cultured cells, and tissues. The first particle delivery system contained a helium propulsion unit (the gun) mounted in a vacuum-controlled chamber. In contrast, the hand-held gene gun does not operate within a chamber. It is completely hand-held, easy, and efficient to use, and it requires minimal space on the laboratory bench top. This chapter describes protocols for using a hand-held gene gun to deliver transformation vectors for overexpression of genes or gene replacement into the macronucleus of Tetrahymena thermophila. The protocols provide helpful information for preparing Tetrahymena for biolistic bombardment, preparation of vector-coated microcarriers, and basic gene gun operating procedures.
弹道式微粒子弹射法被广泛应用于将载体包裹的微粒子递送至微生物、培养细胞和组织中。第一个粒子输送系统包含一个氦气推进装置（枪），安装在一个真空控制室内。相比之下，手动基因枪不在室内运作。它完全由手动操作，使用方便高效，在实验台上所需空间极小。本章描述了使用手动基因枪将转化载体递送至嗜热四膜虫的大核中，以实现基因的过表达或基因替换的方案。这些方案为嗜热四膜虫的生物弹击准备、载体包裹微载体的制备以及基本基因枪操作程序提供了有用信息。
