[The PTD domain of Tat protein enhance GFP protein delivering into myeloma cell SP2/0].
In order to detect the protein delivery mediated by the PTD (protein transduction domain) of TAT Protein, a expression vector, named pT7460-GFP, was constructed by insert the PTD DNA Sequence, followed by a GFP (green fluorescent protein) gene fused in-frame, into the pT7450 vector. The TAT-GFP fusion protein was expressed in the E. coli ER2566. Most of the fusion protein was presented in the inclusion body. The protein was purified by Ni2+ affinity chromatography under denature conditions, then by a Sepharose Q column to remove urea. The soluble denatured protein was added directly to medium containing the Myeloma Cell SP2/0. It came out that the fusion protein could be detected delivered into the cells under fluorescent microscope in a short time.
Li
Zhi-Qi
ZQ
Shanghai Research Center of Biotechnology, Chinese Academy of Sciences, Shanghai 200233, China.
Hu
Xiao-Bo
XB
Yang
Sheng-Li
SL
Gong
Yi
Y
D015696:Gene Products, tat / Q000737:chemistry / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009101:Multiple Myeloma / Q000378:metabolism*; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D015696:Gene Products, tat; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins