[Prokaryotic expression of fusion protein TAT-EDAG and study on its transduction activity].
HIV-TAT protein transduction domain (PTD) is a new kind of peptide that is responsible for transduction of proteins through the plasma membrane with high efficiency. Linked covalently to proteins, peptides, nucleic acid, it could transduce into nearly all kinds of cells and tissues with high efficiency and without any damages. In this study, we constructed the TAT-EDAG and TAT-GFP prokaryotic expression vectors and expressed soluble TAT-EDAG and TAT-GFP fusions in E. coli BL21 (DE3) successfully. By using the Ni-NTA-agrose purification system under the native condition we got the purified fusion proteins whose purification were higher than 90%. After desalting we found the TAT-GFP could transduced successfully into the mouse fibroblast cells and the TAT-EDAG could transduced into HL-60 cells in vitro. It will be useful to amplify the HSCs in vitro in the next step.
Zhou
Sheng-Hua
SH
Institute of Prologue and Veterinary Sciences Academy of Shanxi Agricultural Sciences, Taiyuan 030032, China.
He
Dong-Chang
DC
Zhang
Li-Jun
LJ
Chen
Li-Dong
LD
D000818:Animals; D004926:Escherichia coli / Q000235:genetics; D005347:Fibroblasts / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D051379:Mice; D009687:Nuclear Proteins / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D014161:Transduction, Genetic; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000235:genetics*
C435745:HEMGN protein, human; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D054322:tat Gene Products, Human Immunodeficiency Virus