Clontech 酵母双杂交系统 pGBKT7， 酵母双杂交系统 pGBKT7，Clontech pGBKT7
pGBKT7
产品参数:
Mammalian Selection：TRP1 （HIS3 selection）
载体抗性:
卡那霉素（Kanamycin）
说明书:
质粒图谱:
标签归档：clontech
Clontech pBridge酵母三杂交系统说明书
Clontech pBridge酵母三杂交系统说明书
载体描述:
pBridge expresses two proteins: a DNA-binding domain fusion, and an additional protein. pBridge thus allows establishment of three-hybrid systems when used in combination with an activation domain fusion vector and yeast strains from any of Clontech’s GAL4-based two-hybrid systems, including Matchmaker™ Gold. This vector generates a hybrid protein that contains the sequences for the GAL4 DNA-binding domain （DNA-BD） and the sequence cloned into MCS I. The fusion protein is expressed in yeast host cells from the constitutive ADH1 promoter; transcription is terminated at the ADH1 transcription termination signal. The hybrid protein is targeted to the yeast nucleus by nuclear localization sequences （NLS） that are an intrinsic part of the GAL4 DNA-BD. An additional gene of interest can be cloned into MCS II which is located downstream of an HA epitope and a second NLS. The resulting fusion protein is conditionally expressed from the Met25 promoter in response to methionine levels in the medium; i.e., it is repressed in the presence of 1 mM methionine and expressed in the absence of methionine.
质粒图谱:
酵母单杂交系统Clontech pHis2说明书
酵母单杂交系统 Clontech pHis2说明书
Description
pHIS2 is a reporter vector that can be used in yeast one-hybrid assays to identify and characterize DNA-binding proteins. The vector was specifically designed for use with the BD Matchmaker One-Hybrid Library Construction & Screening Kit （#K1617-1）. It contains a HIS3 nutritional reporter gene, located downstream of a multiple cloning site （MCS） and the minimal promoter of the HIS3 locus （PminHIS3）.Cis-acting DNA sequences, or DNA target elements, can be inserted into the MCS and used as baits to screen GAL4 AD/cDNA fusion libraries for proteins that interact with the target sequence. A protein-DNA （or one-hybrid） interaction can be detected by performing the assay in a yeast strain such as Y187 that is auxotrophic for histidine. Positive one-hybrid interactions drive expression of the HIS3 reporter gene, which enables the host cell to grow on histidine-deficient media.
In the absence of activation, the constitutive HIS3 expression from PminHIS3 is very low. During library screening, the leaky expression of HIS3 is controlled by adding 3-ami,2,4-triazole （3-AT） to the medium. The concentration of 3-AT needed to fully suppress leaky HIS3 expression must be determined empirically for each DNA target element.
pHIS2 can be maintained in both yeast and bacteria. It contains an autonomous replication sequence （ARS4） and TRP1 nutritional marker for replication and selection in yeast （1, 2）; it contains a Col E1 origin and a kanamycin resistance gene （Kanr） for propagation and selection in E. coli. The centromeric sequence CEN6 ensures proper segregation of the plasmid during cell division in yeast （1, 2）.
Use
To use pHIS2 in a one-hybrid assay, clone one or more copies of a cis-acting DNA target sequence into the MCS. Then introduce the plasmid into competent yeast cells using the transformation protocol in the BD Matchmaker Library Construction & Screening Kits User Manual （PT3529-1）. In contrast to the original BD Matchmaker One-Hybrid System, this reporter vector does not need to be integrated into the yeast genome. Instead, it is maintained as an episome throughout the assay. Inserting your target element may alter the level of background HIS3 expression. Therefore, constructs should be tested for background （leaky） HIS3 expression before you start a one-hybrid analysis. Background growth due to leaky HIS3 expression is controlled by adding 3-AT to the selection medium, as described in the User Manual （PT3529-1）.
Location of Features
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Multiple cloning sites: 141
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HIS3 gene: 152811
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Fragment containing the HIS3 3′ UTR & Termination sequence: 8121446
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TRP1 gene: 28553529
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Fragment containing the Col E1 E. coli origin of replication: 41654615
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Kanamycin-resistance gene: 56054811
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CEN6/ARS4 sequences: 62545737
Clontech 代理
Clontech 代理
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部分产品目录
| Brand | Cat. # | Product | Package Size |
| clontech | 636153 | Human Aorta Poly A+ RNA | 5 ug |
| clontech | 636170 | Human Blood, Peripheral Leukocytes Poly A+ RNA | 5 ug |
| clontech | 636113 | Human Heart Poly A+ RNA | 5 ug |
| clontech | 636171 | Human Heart, Auricle Dextra （right） Poly A+ RNA | 5 ug |
| clontech | 636172 | Human Heart, Auricle Sinistra （left） Poly A+ RNA | 5 ug |
| clontech | 636175 | Human Heart, Pericardium Poly A+ RNA | 5 ug |
| clontech | 636173 | Human Heart, Ventricle （left） Poly A+ RNA | 5 ug |
| clontech | 636174 | Human Heart, Ventricle （right） Poly A+ RNA | 5 ug |
| clontech | 636143 | Human Lymph Node Poly A+ RNA | 5 ug |
| clontech | 636121 | Human Spleen Poly A+ RNA | 5 ug |
| clontech | 639537 | SMARTScribe Reverse Transcriptase | 100 Rxns |
| clontech | 634925 | SMARTer PCR cDNA Synthesis Kit | 10 Rxns |
上海金畔生物科技有限公司
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