pMAL-c2x 载体

质粒类型: 大肠杆菌表达载体
表达水平:
启动子: Tac
克隆方法: 多克隆位点,限制性内切酶
载体大小: 6646 bp (查看载体序列)
5' 测序引物及序列: MalE引物: 5'-GGTCGTCAGACTGTCGATGAAGCC-3';
MBP-F: 5'-gatgaagccctgaaagacgcgcag-3'
3' 测序引物及序列: pBad-R:  5'-gatttaatctgtatcagg-3';
M13-F: 5'-TGTAAAACGACGGCCAGT-3'
载体标签: N-MBP, N-Factor Xa
载体抗性: Ampicillin (氨苄青霉素)
备注: 融合表达麦芽糖结合蛋白MBP,蛋白定位于细胞周质
产品编号 产品名称 规格 价格
VT1247 pMAL-c2x 2ug 点击询价

pMAL™系统是一种高效的蛋白融合表达及纯化系统。pMAL载体含有编码麦芽糖结合蛋白(Maltose Binding Protein, MBP)的大肠杆菌malE基因,其下游的多克隆位点便于目的基因插入,表达N端带有MBP的融合蛋白。通过"tac"强启动子和malE翻译起始信号使克隆基因获得高效表达,并进一步利用MBP对麦芽糖的亲和性达到用Amylose柱对融合蛋白的一步亲和纯化。

The system uses the pMAL vectors which are designed so that insertion interrupts a lacZα gene allowing a blue-to-white screen for inserts on X-gal (5). pMAL-c2 series has an exact deletion of the malE signal sequence, resulting in cytoplasmic expression of the fusion protein. pMAL-p2 series contains the normal malE signal sequence, which directs the fusion protein through the cytoplasmic membrane. pMAL-p2 fusion proteins capable of being exported can be purified from the periplasm. Between the malE sequence and the polylinker there is a spacer sequence coding for 10 asparagine residues. This spacer insulates MBP from the protein of interest, increasing the chances that a particular fusion will bind tightly to the amylose resin. The vectors also include a sequence coding for the recognition site of a specific protease. This allows the protein of interest to be cleaved from MBP after purification, without adding any vector-derived residues to the protein (6). For this purpose, the polylinker includes a restriction site superimposed on the sequence coding for the site of the specific protease. This is where the gene of interest is inserted. An EcoR I site in the same reading frame as that of λgt11 and a number of other useful sites are present directly downstream. The vectors also include the M13 origin of DNA replication which allows the production of single-stranded DNA for sequencing and mutagenesis by infecting with M13KO7 helper phage (NEB #N0315S).