Henthorn P, Zervos P, Raducha M, Harris H, Kadesch T
Department of Human Genetics, University of Pennsylvania School of Medicine, Philadelphia 19104-6072.
Proc Natl Acad Sci U S A. 1988 Sep;85(17):6342-6. doi: 10.1073/pnas.85.17.6342.
The human placental alkaline phosphatase gene has been cloned and reintroduced into mammalian cells. When a plasmid carrying the gene under control of the simian virus 40 early promoter (pSV2Apap) is transfected into a variety of different cell types, placental alkaline phosphatase activity can readily be detected by using whole cell suspensions or cell lysates. Alkaline phosphatase activity can also be visualized directly in individual transfected cells by histochemical staining. The gene is appropriate for use as a reporter in studies of gene regulation since its expression is dependent on the presence of exogenous transcription control elements. The overall assay to detect the expression of the gene is quantitative, very rapid, and inexpensive. Cotransfections of cells with pSV2Apap and a related plasmid carrying the bacterial chloramphenicol acetyltransferase gene (pSV2Acat) indicate that transcription of these two genes is detected with roughly the same sensitivity.
人胎盘碱性磷酸酶基因已被克隆并重新导入哺乳动物细胞。当携带在猿猴病毒40早期启动子(pSV2Apap)控制下的该基因的质粒转染到多种不同细胞类型中时,通过使用全细胞悬液或细胞裂解物能够很容易地检测到胎盘碱性磷酸酶活性。碱性磷酸酶活性也可通过组织化学染色在单个转染细胞中直接显现出来。该基因适合用作基因调控研究中的报告基因,因为其表达依赖于外源转录控制元件的存在。检测该基因表达的整体测定是定量的、非常快速且成本低廉的。用pSV2Apap和携带细菌氯霉素乙酰转移酶基因的相关质粒(pSV2Acat)对细胞进行共转染表明,这两个基因的转录检测灵敏度大致相同。