Modification of Enhanced Green Fluorescent Protein for Secretion out of Cells

作者：Roh Ji Yeol; Koo Bon Chul; Kwon Mo Sun; Kim Minjee; Kim Nam Hyung; Kim Teoan^*

来源：Biotechnology and Bioprocess Engineering, 2013, 18(6): 1135-1141.

DOI：10.1007/s12257-013-0333-1

• green fluorescent protein
• secretory protein
• follicle-stimulating hormone
• gene expression
• retrovirus vector

Since its discovery approximately 20 years ago, green fluorescent protein (GFP) has become one of the most widely used reporter proteins. GFP has been used in a variety of living organisms, ranging from E. coli to higher eukaryotes, such as plants and animals. The biggest advantage of using this reporter protein is that it can be used to monitor in vitro and in vivo gene expression. One important limitation, however, is its inability to be secreted out of cells. For this reason, it has been difficult to directly measure the expression level of the regulatory sequence of a gene of interest quantitatively. To overcome this drawback, we have modified the enhanced green fluorescent protein gene (EGFP), a derivative of GFP, by adding a signal peptide sequence that encodes a rat follicle-stimulating hormone (FSH) beta-subunit upstream of EGFP. Following the expression of this modified gene in several cell types, we have found efficient secretion of EGFP. Consequently, with the secreted protein, we could easily quantify the gene expression level with high reliability. Therefore, the use of our modified EGFP expression cassette would greatly facilitate the evaluation of regulatory sequences, such as promoters and enhancers. Further, it will also be very helpful in the study of transgenic livestock intended to use as bioreactors for mass production of pharmaceuticals.

• 出版日期2013-11