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pAbAi
    pAbAi
  • 平臺(tái)編號(hào):bio-109376
  • 質(zhì)粒信息: 宿主:DH5α,BL21
  • 規(guī)格:2ml甘油菌 20ul質(zhì)粒
  • 用途:酵母系列,酵母單雜交載體,配套pGADT7,低拷貝載體
  • 服務(wù)費(fèi)用:
    加載中……
  • 訂購(gòu)
  • 注意事項(xiàng):僅用于科學(xué)研究或者工業(yè)應(yīng)用等非醫(yī)療目的不可用于人類或動(dòng)物的臨床診斷或治療,非藥用,非食用(產(chǎn)品信息以出庫(kù)為準(zhǔn))

啟動(dòng)子 URA3
復(fù)制子 pUC
質(zhì)粒分類 酵母系列質(zhì)粒;酵母雜交質(zhì)粒;單雜交類質(zhì)粒
質(zhì)粒大小 4894bp
原核抗性 Amp
篩選標(biāo)記 URA3
克隆菌株 DH5a
培養(yǎng)條件 30℃,YPDA,有氧
表達(dá)宿主 Y1Hgold等酵母菌
載體宿主 酵母菌
載體用途 雜交
原核抗性 Amp
真核抗性 URA3
5'測(cè)序引物 pABAI-F(GTTCCTTATATGTAGCTTTCGACA)
3'測(cè)序引物 pABAI-R(CCATCTCGAAAAAGGGTTTGCC)
備注 低拷貝質(zhì)粒
質(zhì)粒簡(jiǎn)介 pABAI質(zhì)粒是一種單雜交酵母報(bào)告質(zhì)粒,可以識(shí)別和分析蛋白質(zhì)與DNA之間的相互作用,而不僅僅是證明蛋白之間的相互作用。這個(gè)質(zhì)粒具有AbA抗性因而具有很強(qiáng)的篩選能力,可極大地降低背景水平。
          pABAI is a yeast reporter vector that can be used in one-hybrid assays to identify and characterize DNA-binding proteins. The vector, which was specifically designed for use with the Matchmaker Gold Yeast One-Hybrid Library Screening System, contains a multiple cloning site upstream of the yeast iso-1-cytochrome C minimal promoter(not shown) and the AUR1-C gene, an antibiotic resistance gene that confers resistance to Aureobasidin A. A cis-acting element (i.e., a target sequence) can be inserted into the MCS and used as bait to screen GAL4 AD/cDNA fusion libraries for proteins that interact with the target sequence. Positive protein-DNA (i.e.,one-hybrid) interactions drive the expression of AUR1-C. As a result, one-hybrid interactions can be detected by selecting for yeast that are resistant to AbA.
          pABAI cannot be propagated episomally in yeast; it can only be stablymaintained through integration into the host genome. Integration is accomplished via homologous recombination between the vector’s URA3 gene and the ura3-52 locus of the yeast strain provided in theMatchmaker Gold Yeast One-Hybrid System. URA3 is a nutritional marker that can also be used for selection of recombinant yeast. The vector also contains a Col E1 origin of replication and an ampicillin resistance gene (Ampr) for propagation and selection in E.coli.
          To use pAbAi in a one-hybrid assay, clone one or more copies of a cis-acting element into the MCS. To facilitate recombination of the vector with the host’s ura3-52 locus, linearize the vector with either BbsI or BstBI, then introduce the linearized vector into competent yeast cells using the protocols in the Matchmaker Gold Yeast One-Hybrid Library Screening System User Manual.
          Insertion of your target sequence may alter the basal expression of AUR1-C. Therefore, before starting a onehybrid analysis, basal levels of AUR1-C should be determined as described in the User Manual.
          •    Suitable host strains: DH5α and other general purpose strains.
          •    Selectable marker: plasmid confers resistance to ampicillin (100 µg/ml) to E. coli hosts.
          •    Copy number: low
           

 

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