David J C, Loir M, Lefresne J, Thiebaud P, Signoret J
Laboratoire de Biochimie du Développement, L.A. No 256, C.N.R.S., Université de Rennes I, Campus de Beaulieu, 35042, Rennes Cedex, France.
Laboratoire de Physiologie des Poissons, I.N.R.A., Université de Rennes I, Campus de Beaulieu, 35042, Rennes Cedex, France.
Rouxs Arch Dev Biol. 1986 Apr;195(3):186-192. doi: 10.1007/BF02439437.
During animal development and gametogenesis two DNA ligases are found and successively expressed. In this study the two DNA ligases present in the axolotl egg and the two ligases present during ram sperm cell maturation were distinguished by biochemical and immunological methods. The expression of the genes for the heavy and light ram DNA ligases has been studied using transplantation of spermatid and sperm nuclei in axolotl eggs. We found that ram DNA ligases were expressed in axolotl egg cytoplasm. The exclusion phenomenon between the heavy and light form of DNA ligase is species-specific and involves a cytoplasmic mediator. In the transplanted ram germ cell nuclei the heavy ram DNA ligase expression was found to be sensitive to inhibitors of transcription while the light one was not. When mRNA was used, no exclusion process was observed and both the heavy and light enzyme expression were sensitive to cycloheximide and not to aamanitin. These results are discussed in terms of the possible stability of the gene-regulated state following nuclear transfer.
在动物发育和配子发生过程中,会发现两种DNA连接酶并相继表达。在本研究中,通过生化和免疫学方法区分了蝾螈卵中存在的两种DNA连接酶以及公羊精子细胞成熟过程中存在的两种连接酶。利用精细胞和精核移植到蝾螈卵中的方法,研究了公羊重链和轻链DNA连接酶基因的表达。我们发现公羊DNA连接酶在蝾螈卵细胞质中表达。DNA连接酶重链和轻链形式之间的排斥现象具有物种特异性,且涉及一种细胞质介质。在移植的公羊生殖细胞核中,发现重链公羊DNA连接酶的表达对转录抑制剂敏感,而轻链则不敏感。当使用mRNA时,未观察到排斥过程,重链和轻链酶的表达均对环己酰亚胺敏感,而对放线菌素不敏感。根据核移植后基因调控状态的可能稳定性对这些结果进行了讨论。