Neale Matthew J, Keeney Scott
MRC Genome Damage and Stability Centre, University of Sussex, Falmer, UK.
Methods Mol Biol. 2009;557:183-95. doi: 10.1007/978-1-59745-527-5_12.
During meiosis Spo11 catalyzes the formation of DNA double-strand breaks, becoming covalently attached to the 5' ends on both sides of the break during this process. Spo11 is removed from the DSB by single-stranded endonucleolytic cleavage flanking the DSB, liberating a short-lived species consisting of Spo11 protein covalently linked to a short oligonucleotide. The method presented here details how to detect these Spo11-oligo complexes in extracts made from meiotic yeast cells.
在减数分裂过程中，Spo11催化DNA双链断裂的形成，并在此过程中与断裂两侧的5'端共价连接。通过双链断裂侧翼的单链内切核酸酶切割，Spo11从双链断裂处被移除，释放出一种由Spo11蛋白与短寡核苷酸共价连接组成的短暂存在的物质。本文介绍的方法详细说明了如何在减数分裂酵母细胞提取物中检测这些Spo11-寡核苷酸复合物。
