Molecular analysis of gene conversion in spermatids from transgenic mice.
Molecular analysis of gene conversion in spermatids from transgenic mice.
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转基因小鼠精子细胞基因转化的分子分析。
DOI：
10.1016/s0378-1119(97)00416-2
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发表时间：
1997
期刊：
影响因子：
3.5
通讯作者：
Schimenti,JC
中科院分区：
文献类型：
--
作者：
Hanneman,WH;Schimenti,KJ;Schimenti,JC
由于不能回收减数分裂的所有产物，对哺乳动物基因转换的机制和性质的研究受到很大限制。此外，由于缺乏可见的标记来检测基因转换，特别是当事件罕见时，这一过程的研究受到阻碍。在先前的工作中，我们开发了用于检测和定量小鼠种系中的基因转换事件的转基因系统（Murti，J.R.，Bumbulis，M.，Schimenti，J.C.，1992.转基因小鼠中的高频率生殖系基因转换。摩尔Cell. 12，2545-2552），其可以用作重组化学物质的测定（Murti，J.R，Schimenti，K. J.，Schimenti，J.C.，1994.一种用于遗传毒性检测的重组转基因小鼠系统。穆塔Res. 307，583-595）。两个互补缺陷lacZ基因之间的特定染色体内基因转换事件导致精子细胞中β-半乳糖苷酶的产生，从而能够测量转换频率。在这里，我们报告说，抗癌药物，顺铂，增加在这些转基因小鼠减数分裂阶段细胞的基因转换。此外，开发了一种用于直接分子分析单个或合并的lacZ阳性精子细胞中的转基因转换事件的方法。鉴定经历了罕见的基因转换事件的配子的能力，随后是重组基因的分子扩增，应该使得有可能比以前更详细地研究哺乳动物中的遗传重组机制。
Investigations into the mechanisms and properties of gene conversion in mammals are greatly restricted by the inability to recover all the products of a meiosis. Additionally, the study of this process has been hampered by the lack of visible markers to detect gene conversion, especially when the events are rare. In previous work, we developed a transgenic system for detection and quantitation of gene conversion events in the germline of mice (Murti, J.R., Bumbulis, M., Schimenti, J.C., 1992. High frequency germline gene conversion in transgenic mice. Mol. Cell. Biol. 12, 2545–2552) that could be exploited as an assay for recombinogenic chemicals (Murti, J.R, Schimenti, K.J., Schimenti, J.C., 1994. A recombination-based transgenic mouse system for genotoxicity testing. Mutat. Res. 307, 583–595). A specific intrachromosomal gene conversion event between two complementarily defective lacZ genes resulted in the production of β-galactosidase in spermatids, enabling a measurement of conversion frequency. Here, we report that the anticancer drug, cisplatin, increased gene conversion in meiotic stage cells in these transgenic mice. Furthermore, a method was developed for direct molecular analysis of transgene conversion events in single or pooled lacZ-positive spermatids. The ability to identify gametes that have undergone a rare gene conversion event, followed by molecular amplification of the recombinant gene, should make it possible to investigate the mechanisms of genetic recombination in mammals in greater detail than previously possible.