pMAL-c4x

质粒类型：大肠杆菌表达载体

表达水平：高

启动子：Tac

克隆方法：多克隆位点，限制性内切酶

载体大小：6645 bp

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

The vector pMAL-c4X is designed to produce maltose-binding protein (MBP) fusions, where the protein of interest can be cleaved from MBP with the specific protease Factor Xa.
MBP fusions made with this vector are expressed cytoplasmically. The MBP has been engineered for tighter binding to amylose resin. 
A gene or open reading frame is inserted into a restriction site of the vector polylinker, in the same translational reading frame as the malE gene (encoding maltose-binding protein). Insertion of the DNA fragment interrupts the malE-lacZα fusion pre-existing on the vector, affording a screen for inserts on the proper indicator plates. The fusion protein thus produced can be purified by amylose affinity chromatography. The sequence coding for the four amino acids Ile-Glu-Gly-Arg is present just upstream of the XmnI site. This allows the protein of interest to be cleaved from maltose-binding protein with the specific protease Factor Xa. Fragments inserted in the XmnI site (cleaves GAAGG↓ATTTC) will produce a fusion protein that, after Factor Xa cleavage, contains no vector-derived residues on the protein of interest.

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