High expression of synthetic human interferon-gamma cDNA in E. coli.
Human interferon-gamma (IFN-gamma) cDNA was synthesized, and it makes the usage of favorable codons in E. coli. The authors got 9 different expression plasmids which contain the synthetic IFN-gamma-cDNA and have different spaces between SD sequence and ATG. The free energies G0f298 in the formation of stable secondary structure in the translation initiation region (TIR) are different in various expression plasmids. One of them, pLY4-gamma 5, can highly yield INF-gamma which will be about 60%-80% of the total bacterial proteins, such a high expression was hardly noted in literature. The reasons of high expression in this work are optimal spaces between SD and ATC, favorable delta G0f298, favourable condons usage for E. coli.
Wang
Z
Z
Shanghai Institute of Biochemistry, Chinese Academy of Sciences, China.
Yang
D
D
Wang
Q
Q
Li
B
B
Lü
Z
Z
Yu
J
J
Zheng
H
H
Fan
P
P
Tang
J
J
Qian
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D003062:Codon; D018076:DNA, Complementary / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins
D003062:Codon; D018076:DNA, Complementary; D011994:Recombinant Proteins; D007371:Interferon-gamma