The use of lacZ gene fusions in the studies of mammalian development: developmental regulation of mammalian homeobox genes in the CNS.

Journal de physiologie Pub Date : 1990-01-01
J Zakany, C K Tuggle, C M Nguyen-Huu
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Abstract

1. Mammalian cells can produce bacterial beta-galactosidase when they carry the lacZ gene under the control of mammalian regulatory elements. The single cell resolution of the beta-galactosidase histochemical detection method makes this molecule an excellent marker in studies of development at the cellular and molecular level. Different lacZ fusion genes can be engineered to study the histological diversification of cell lineages, the developmental regulation of isolated genes, or to recognize and clone genes with new expression profile. 2. Transgenic mice carrying lacZ gene fusions provide information on the cell type, developmental stage and spatial specificity of cis-acting regulatory regions linked to a mammalian homeobox gene. We describe our strategy for designing the gene fusions. 3. The scord region of the Hox 1.3 gene is sufficient to determine spatially restricted expression of a heterologous protein in the midgestational spinal cord. We propose to use this region to alter the expression pattern of other homeobox gene products. Developmental alterations due to a variant expression pattern would point to the function of the misexpressed gene.

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lacZ基因融合在哺乳动物发育研究中的应用:哺乳动物中枢神经系统同源盒基因的发育调控。
1. 哺乳动物细胞携带lacZ基因,在哺乳动物调控元件的控制下,可以产生细菌β -半乳糖苷酶。-半乳糖苷酶组织化学检测方法的单细胞分辨率使该分子在细胞和分子水平上的发育研究中成为一个很好的标记物。不同的lacZ融合基因可以用于研究细胞系的组织学多样化、分离基因的发育调控,或识别和克隆具有新表达谱的基因。2. 携带lacZ基因融合物的转基因小鼠提供了与哺乳动物同源盒基因相关的顺式调控区域的细胞类型、发育阶段和空间特异性的信息。我们描述了我们设计基因融合的策略。3.Hox 1.3基因的scord区域足以确定孕中期脊髓中异源蛋白的空间限制性表达。我们建议利用这个区域来改变其他同源盒基因产物的表达模式。由于变异表达模式导致的发育改变将指向错误表达基因的功能。
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