Expression of a mouse metallothionein-Escherichia coli beta-galactosidase fusion gene (MT-beta gal) in early mouse embryos.
Expression of a mouse metallothionein-Escherichia coli beta-galactosidase fusion gene (MT-beta gal) in early mouse embryos.
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DOI：
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发表时间：
1989
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。以5-溴-4-氯-3-吲哚基β-D-吡喃半乳糖苷（X-Gal）为底物，采用定性组织化学方法检测不同着床前阶段lacZ的表达。我们观察到染色指示外源性β-半乳糖苷酶的活性在5-17%的DNA注射胚胎植入前阶段用硫酸锌处理16-24小时后测定。因此，lacZ可用作小鼠胚胎发生早期启动子功能的指示基因，并且将MT-1启动子掺入融合基因中可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 for Escherichia coli beta-galactosidase (lacZ), into the pronuclei of one-cell mouse embryos. A qualitative histochemical assay, with 5-bromo-4-chloro-3-indolyl beta-D-galactopyranoside (X-Gal) as a substrate, was used to detect expression of lacZ at several preimplantation stages. We observed staining indicative of exogenous beta-galactosidase activity in 5-17% of DNA-injected embryos assayed at preimplantation stages after 16-24 h treatment with ZnSO4. Thus, lacZ can 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.