Liposome membrane itself can affect gene expression in the Escherichia coli cell-free translation system.
A possible role of a model biomembrane, liposome, in gene expression was investigated by using the cell-free translation system. A reporter protein, green fluorescence protein (GFP), was expressed in vitro with and without liposome prepared with 1-palmitoyl-2-oleoyl- sn-glycero-3-phosphatidyl chorine (POPC) and cholesterol (Ch) (5.7 mM lipid concentration). In the presence of POPC/Ch liposome, the fluorescence intensity of produced GFP was found to be 1.67 times higher than that in the control after 18 h of expression. The results of the SDS-PAGE analysis also show the above promotion effect of the liposome on the net expression of the GFP gene (1.58 times more). The amounts of mRNA were found to be promoted to 1.29 times higher than those in the control. The differences among mRNA, net expression of the GFP gene, and GFP fluorescence indicate that the enhanced GFP expression in the presence of POPC/Ch liposome could primarily affect the transcription and translation of the GFP gene among the possible steps of gene expression. The variation of in vitro gene expression with various liposomes also shows that the biomembrane could act as a modulator to split the genotype and phenotype in a biological cell.
Bui
Huong Thi
HT
Department of Chemical Science and Engineering, Graduate School of Engineering Science, Osaka University, Toyonaka, Osaka, Japan.
Umakoshi
Hiroshi
H
Ngo
Kien Xuan
KX
Nishida
Masato
M
Shimanouchi
Toshinori
T
Kuboi
Ryoichi
R