2、 Vector载体简介The vector pMAL‐p5X 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 include an N‐terminal signal sequ
3、ence, so the fusion protein is directed to the periplasm. 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 fra
4、me as the malE gene (encoding maltose‐binding protein).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 al
5、lows 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‐de
6、rived residues on the protein of interest.pMAL‐p5x载体序列ORIGIN1CCGACACCATCGAATGGTGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGA61GTCAATTCAGGGTGGTGAATGTGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCG121GTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAA181CG
