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原核显微注射后SV40-lacZ基因在小鼠植入前胚胎中的表达。

Expression of SV40-lacZ gene in mouse preimplantation embryos after pronuclear microinjection.

作者信息

Takeda S, Toyoda Y

机构信息

Department of Animal Pathology, University of Tokyo, Japan.

出版信息

Mol Reprod Dev. 1991 Oct;30(2):90-4. doi: 10.1002/mrd.1080300203.

Abstract

In order to study the expression of an exogenous gene in developing mouse embryos during the preimplantation period, DNA carrying the SV40 early promoter fused with the Escherichia coli beta-galactosidase gene (lacZ) was microinjected into the pronucleus of fertilized mouse eggs. Expression of lacZ gene was detected by staining embryos with 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) as a substrate at pH 7.2. The embryos expressing the lacZ gene showed various intensities of blue staining, all showing a mosaic pattern. The exogenous gene was expressed from the 4-cell stage until the blastocyst stage. The proportion of embryos expressing the lacZ gene was maximal (38%) at the morula stage, and the expression was dependent on the presence of the SV40 promoter.

摘要

为了研究植入前阶段发育中的小鼠胚胎中外源基因的表达情况,将携带与大肠杆菌β-半乳糖苷酶基因(lacZ)融合的SV40早期启动子的DNA显微注射到受精小鼠卵的原核中。以5-溴-4-氯-3-吲哚基-β-D-吡喃半乳糖苷(X-gal)为底物,在pH 7.2条件下对胚胎进行染色,检测lacZ基因的表达。表达lacZ基因的胚胎呈现出不同强度的蓝色染色,均表现为镶嵌模式。外源基因从4细胞阶段一直表达至囊胚阶段。在桑椹胚阶段,表达lacZ基因的胚胎比例最高(38%),且该表达依赖于SV40启动子的存在。

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