pSV-?-Galactosidase Control Vector

• 产品编号：E1081      品牌：promega       原厂货号：E1081
• 产品分类：克隆 > 克隆载体和克隆试剂盒 > 不依赖DNA连接酶的克隆试剂盒 > 其它克隆载体
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其它组分：

描述：

pSV-β-Galactosidase Control Vector(pSV-β- 半乳糖苷酶对照载体) 是监控哺乳动物细胞转染效率的阳性对照载体。其中，SV40早期启动子(SV40 early promoter) 和增强子驱动lac Z 基因的转录；lac Z 基因编码β- 半乳糖苷酶。pSV-β- 半乳糖苷酶对照载体既可单独转染，也可与您的目的DNA 共转染。β- 半乳糖苷酶是一种很好的报告基因酶，可以用分光光度法、荧光法或化学发光法快速直接地在细胞提取物中检测。使用X-Gal 作底物，这个报告基因酶也被广泛用于原位组化分析。

pSV-β- 半乳糖苷酶对照载体可与您的目的DNA 共转染。例如，如果将该载体与萤火虫萤光素酶基因载体(pGL3 载体) 进行共转染，能够同时检测细胞提取物中的萤光素酶和β- 半乳糖苷酶的活性。在这种情况下，pSV-β- 半乳糖苷酶载体作为瞬时表达检测的内对照。实验也应检测阴性对照提取物，用于判定细胞培养物中有无内源性β- 半乳糖苷酶，该阴性对照提取物由假转染的细胞制备而来。此外，共转染氯霉素乙酰转移酶报告基因载体(pCAT^® 3 载体)，就能够检测CAT 和β- 半乳糖苷酶的活性。pSV-β- 半乳糖苷酶载体是由pRSV-β-Gal 改造而来，将SV40 和pUC18 序列取代了RSV 和pBR322 序列。pSV-β- 半乳糖苷酶载体可在E. coli 内表达pSV-β- 半乳糖苷酶，因为lac Z 基因的上游为E. coli gpt 启动子。带有pSV-β-半乳糖苷酶载体的E. coli 克隆在含有X-Gal 的培养基上呈蓝色。

原厂资料：

The pSV-β-Galactosidase Control Vector is designed as a positive control vector to monitor transfection efficiencies of mammalian cells. The SV40 early promoter and enhancer drive transcription of the bacterial lacZ gene, which in turn, is translated into the β-galactosidase enzyme. β-galactosidase is an excellent reporter enzyme that can be assayed quickly and directly in cell extracts using spectrophotometric, fluorescent or chemiluminescent assays. This reporter enzyme also is widely used for in situ histochemical analysis using the substrate X-Gal.
The pSV-β-Galactosidase Control Vector can be co-transfected with your DNA of interest. For example, co-transfection with firefly luciferase gene vectors (pGL3 Vectors) provide cell extracts that can be assayed for both luciferase and β-galactosidase activities. In this manner, the pSV-β-Galactosidase Vector acts as an internal control for transient expression assays. A negative control extract, prepared from mock-transfected cells, also should be assayed for the presence of endogenous β-galactosidase activity in cultured cells. In addition, co-transfection with chloramphenicol acetyltransferse reporter gene vectors (pCAT®3 Vectors) permits assaying both CAT and β-galactosidase activities.
The pSV-β-Galactosidase Vector is a modification of pRSV-βGAL with SV40 and pUC18 sequences substituted for RSV and pBR322 sequences. The pSV-β-Galactosidase Vector will express β-galactosidase in E. coli due to the presence of the E. coli gpt promoter located upstream of the lacZ gene. Colonies of E. coli containing the pSV-β-Galactosidase Vector will appear blue when plated on media containing X-gal.

注意事项：

储存于-20°C。