pMAL-c5x載體基本信息
載體名稱: | pMAL-c5x |
---|---|
質(zhì)粒類型: | 大腸桿菌表達(dá)載體 |
高拷貝/低拷貝: | -- |
啟動(dòng)子: | -- |
克隆方法: | 多克隆位點(diǎn),限制性內(nèi)切酶 |
載體大小: | 5677bp |
5' 測(cè)序引物及序列: | -- |
3' 測(cè)序引物及序列: | -- |
載體標(biāo)簽: | -- |
載體抗性: | Ampicillin |
篩選標(biāo)記: | -- |
備注: | -- |
產(chǎn)品目錄號(hào): | -- |
穩(wěn)定性: | -- |
組成型: | -- |
病毒/非病毒: | -- |
The vector pMAL-c5X 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 (NEB #P8010).
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). 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.