Expression of a mouse metallothionein-Escherichia coli β-galactosidase fusion gene (MT-βgal) in early mouse embryos☆
Expression of a mouse metallothionein-Escherichia coli β-galactosidase fusion gene (MT-βgal) in early mouse embryos☆
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DOI：
10.1016/0014-4827(89)90392-3
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发表时间：
1989-08
影响因子：
3.7
通讯作者：
M. Stevens;J. Meneses;R. Pedersen
中科院分区：
文献类型：
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作者：
M. Stevens;J. Meneses;R. Pedersen
我们将含有诱导型小鼠金属硫蛋白-I (MT-I) 启动子的 DNA 显微注射到单细胞小鼠胚胎的原核中，该启动子与大肠杆菌β-半乳糖苷酶 (lacZ) 的结构基因偶联。采用定性组织化学测定，以 5-溴-4-氯-3-吲哚基β-d-吡喃半乳糖苷 (X-Gal) 作为底物，用于检测 lacZ 在几个植入前阶段的表达。我们在植入前阶段用 ZnSO4 处理 16-24 小时后，在 5-17% 的 DNA 注射胚胎中观察到了表明外源 β-半乳糖苷酶活性的染色结果。因此，lacZ可以作为小鼠早期胚胎发生过程中启动子功能的指示基因，将MT-I启动子掺入融合基因可以成为控制植入前小鼠胚胎中外源基因表达的有用手段。
We have microinjected DNA containing the inducible mouse metallothionein-I (MT-I) promoter, coupled to the structural gene forEscherichia coliβ-galactosidase (lacZ), into the pronuclei of one-cell mouse embryos. A qualitative histochemical assay, with 5-bromo-4-chloro-3-indolylβ-d-galactopyranoside (X-Gal) as a substrate, was used to detect expression oflacZat several preimplantation stages. We observed staining indicative of exogenous β-galactosidase activity in 5–17% of DNA-injected embryos assayed at preimplantation stages after 16–24 h treatment with ZnSO4. Thus,lacZcan be used as an indicator gene for promoter function during early mouse embryogenesis, and the incorporation of the MT-I promoter into fusion genes can be a useful means of controlling the expression of exogenous genes in preimplantation mouse embryos.