Winkler C, Vielkind J R, Schartl M
Genzentrum, Max-Planck-Institut für Biochemie, Martinsried, FRG.
Mol Gen Genet. 1991 Apr;226(1-2):129-40. doi: 10.1007/BF00273596.
Species of small fish are becoming useful tools for studies on vertebrate development. We have investigated the developing embryo of the Japanese medaka for its application as a transient expression system for the in vivo analysis of gene regulation and function. The temporal and spatial expression patterns of bacterial chloramphenicol acetyltransferase and galactosidase reporter genes injected in supercoiled plasmid form into the cytoplasm of one cell of the two-cell stage embryo was promoter-specific. The transient expression was found to be mosaic within the tissue and organs reflecting the unequal distribution of extrachromosomal foreign DNA and the intensive cell mixing movements that occur in fish embryogenesis. The expression data are consistent with data on DNA fate. Foreign DNA persisted during embryogenesis and was still detectable in some 3- and 9-month-old adult fish; it was found in high molecular weight form as well as in circular plasmid conformations. The DNA was replicated during early and late embryogenesis. Our data indicate that the developing medaka embryo is a powerful in vivo assay system for studies of gene regulation and function.
小型鱼类正成为脊椎动物发育研究的有用工具。我们研究了日本青鳉发育中的胚胎,将其用作瞬时表达系统,用于体内基因调控和功能分析。以超螺旋质粒形式注入两细胞期胚胎一个细胞的细胞质中的细菌氯霉素乙酰转移酶和半乳糖苷酶报告基因的时空表达模式具有启动子特异性。发现瞬时表达在组织和器官内呈镶嵌性,这反映了染色体外外源DNA的不均匀分布以及鱼类胚胎发生过程中发生的强烈细胞混合运动。表达数据与DNA命运的数据一致。外源DNA在胚胎发生过程中持续存在,在一些3个月和9个月大的成年鱼中仍可检测到;它以高分子量形式以及环状质粒构象存在。DNA在胚胎发生的早期和晚期进行复制。我们的数据表明,发育中的青鳉胚胎是研究基因调控和功能的强大体内检测系统。