生物素?；亟M蛋白表達(dá)系統(tǒng) 哺乳動物表達(dá)載體pcDNA6/BioEase表達(dá)生物素酰化的融合蛋白，提供了一種簡便、高效的檢測和純化手段。BioEase標(biāo)簽是含72個氨基酸殘基的肽段，體內(nèi)的生物素連接酶系統(tǒng)（ Biotin Protein Ligase），使這段肽鏈上特定的賴氨酸殘基生物素?；?。生物素?；娜诤系鞍卓梢杂面溍褂H和素瓊脂糖珠進(jìn)行純化。
The pcDNA6-BioEase Gateway Biotinylation System provides an easy, efficient method for expressing, purifying, and detecting biotinylated recombinant proteins. The pcDNA6/BioEase-DEST vector (Figure 1) incorporates a 72 amino acid sequence from K. pneumoniae that directs in vivo biotinylation of a specific lysine residue. This K. pneumoniae tag is recognized and efficiently biotinylated in vivo by a native mammalian system similar to the enzyme Biotin Protein Ligase (BPL) in E. coli(1). Proteins produced from the pcDNA6/BioEase-DEST vector are expressed as fusion proteins to this sequence. Biotinylated fusion proteins can be purified by streptavidin agarose, detected on western blots using streptavidin-HRP or streptavidin-AP conjugates, or visualized in mammalian cells by immunofluorescence with a streptavidin-FITC conjugate (Figure 2). In addition to the 72 amino acid K. pneumoniae sequence, the pcDNA6/BioEase-DEST vector includes the following features:
attR sites for fast and efficient recombination with any attL-flanked Gateway entry vector
The CMV promoter for high-level expression of the recombinant fusion protein
An enterokinase cleavage site for efficient removal of the N-terminal BioEase fusion tag
Gateway哺乳動物表達(dá)載體選擇指南：
pDEST12.2 不帶標(biāo)簽
pDEST26 N-His標(biāo)簽 G418篩選
pDEST27 N-GST標(biāo)簽 G418篩選
pDEST40 C-His標(biāo)簽、C-V5標(biāo)簽 G418篩選
pcDNA-DEST40 C-His標(biāo)簽、C-V5標(biāo)簽 G418篩選
pcDNA-DEST47 C-綠色熒光蛋白 G418篩選
pcDNA-DEST53 N-綠色熒光蛋白 G418篩選
pcDNA3.2/V5-DEST C-V5標(biāo)簽 G418篩選
pcDNA6.2/V5-DEST C-V5標(biāo)簽 Blasticidin S篩選
pcDNA6.2/N-YFP-DEST N-黃色熒光蛋白 Blasticidin S篩選
pcDNA6.2/N-EmGFP-DEST N-綠色熒光蛋白 Blasticidin S篩選
pcDNA6.2/C-YFP-DEST C-黃色熒光蛋白 Blasticidin S篩選
pcDNA6.2/C-EmGFP-DEST C-綠色熒光蛋白 Blasticidin S篩選
pcDNA6.2/BioEase-DEST N-生物素 Blasticidin S篩選
