■ 基本信息
启动子: | URA3 |
复制子: | ColE1 ori |
质粒分类: | 酵母系列质粒；酵母杂交质粒；单杂交类质粒 |
质粒大小: | 4894bp |
原核抗性: | Amp |
真核抗性: | URA3 |
克隆菌株: | DH5a |
培养条件: | 37度 |
表达宿主: | Y1Hgold等酵母菌 |
培养条件: | 30℃，YPDA，有氧 |
5'测序引物: | pABAI-F（GTTCCTTATATGTAGCTTTCGACA） |
3'测序引物:
| pABAI-R（CCATCTCGAAAAAGGGTTTGCC）
|
备注: | 低拷贝质粒 |
■ 质粒属性
质粒宿主: | 酵母菌 |
质粒用途: | 杂交 |
片段类型: |
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片段物种: |
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原核抗性: | Amp |
真核抗性: | URA3
|
荧光标记: |
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■ 质粒简介
pABAI质粒是一种单杂交酵母报告质粒，可以识别和分析蛋白质与DNA之间的相互作用，而不仅仅是证明蛋白之间的相互作用。这个质粒具有AbA抗性因而具有很强的筛选能力，可极大地降低背景水平。
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
■ 质粒图谱
■ 质粒序列
质粒序列请下载：
ZK969pABAI酵母单杂交质粒.txt
