Cervera Amelia, de la Peña Marcos
Instituto de Biología Molecular y Celular de Plantas (IBMCP), Consejo Superior de Investigaciones Científicas-Universitat Politècnica de València (CSIS-UPV), Valencia, Spain.
Methods Mol Biol. 2021;2167:27-44. doi: 10.1007/978-1-0716-0716-9_3.
Retrozymes are a novel family of non-autonomous retrotransposable elements that contain hammerhead ribozyme motifs. These retroelements are found widespread in eukaryotic genomes, with active copies present in many species, which rely on other autonomous transposons for mobilization. Contrary to other retrotransposons, transcription of retrozymes in vivo leads to the formation and accumulation of circular RNAs, which can be readily detected by RNA blotting. In this chapter, we describe the procedures needed to carry out the cloning of genomic retrozymes, and to detect by northern blot their circular RNA retrotransposition intermediates.
逆转录酶是一类新型的非自主逆转座元件家族，其包含锤头状核酶基序。这些逆转元件广泛存在于真核生物基因组中，在许多物种中都有活跃的拷贝，它们依赖于其他自主转座子进行移动。与其他逆转座子不同，逆转录酶在体内的转录会导致环状RNA的形成和积累，通过RNA印迹法可以很容易地检测到这些环状RNA。在本章中，我们描述了进行基因组逆转录酶克隆以及通过Northern印迹法检测其环状RNA逆转座中间产物所需的步骤。
