Soluble expression of recomb inant cMyc, Klf4, Oct4, and Sox2 proteins in bacteria and transduction into living cells

Abstract

The results in the present study indicate the four important reprogramming proteins, cMyc, Klf4, Oct4, and Sox2, can be produced in soluble format in bacteria with low cost. Our new method thus might be expected to greatly contribute to the future study of iPSCs.

Expression of Recombinant Human Octamer-Binding Transcription Factor 4 in Rice Suspension Cells

“…Recombinant Oct4 proteins were produced in mammalian cells [ 18 ], insect cells [ 19 ], yeast [ 20 ], and E. coli . [ 21 ]. Although some reports do evaluate their protein yields, various units were used.…”

“…Although some reports do evaluate their protein yields, various units were used. The yield from previous studies in insect cells and yeast showed that the recombinant Oct4 yield was 6.1 mg/L [ 19 ] and 210 mg/L [ 21 ], respectively. Our present study indicates that the yield of rhOct4 was approximately 0.41% of the total soluble proteins.…”

“…To make the αAmy3 promoter–signal peptide–Oct4 fusion construct, the Gateway-compatible binary T-DNA destination vector, pAAmy3Dst [ 21 ], was used. The 1083-bp Oct4 cDNA fragment was amplified by RT-PCR using the forward primer (5′-CACCATGGCGGGACACCTGGCTTC-3′) and the reverse primer (5′-TCAGTTTGAATGCATGGG-3′).…”

“…To derive the recombinant Oct4 for iPSCs formation, several protein expression platforms have expressed recombinant human Oct4, such as mammalian cells [ 18 ], insect cells [ 19 ], the yeast Pichia pastoris [ 20 ], and Escherichia coli [ 21 ], but some problems have been encountered. In mammalian expression systems, recombinant Oct4 proteins are limited by low yields, cumbersome manipulations, and high culture costs [ 18 ].…”

“…Production of Oct4 in P. pastoris is strongly induced by methanol [ 20 ], but there are concerns about the toxic and inflammatory nature of methanol. In E. coli , recombinant Oct4 proteins are aggregated at inclusion bodies, hence additional denaturation and recovery steps are required during protein purification [ 21 ].…”