A high-level prokaryotic expression system: synthesis of human interleukin 1 alpha and its receptor antagonist.
Synthetic intronless genes, coding for human interleukin 1 alpha (IL 1 alpha) and interleukin 1 receptor antagonist (IL1ra), have been expressed efficiently in a specially designed prokaryotic vector, pGMCE (a pGEM1 derivative), where the target gene forms the second part of a two-cistron system. The first part of the system is a translation enhancer-containing mini-cistron, whose termination codon overlaps the start codon of the target gene. In the case of the IL1 alpha gene, the high expression level is largely due to the direct efficient translation initiation at the second cistron, whereas with the IL1ra gene in the same system, the proximal translation initiation region (TIR) provides a high level of coupled expression of the target gene. Thus, pGMCE is a potentially versatile vector for direct prokaryotic expression.
Birikh
K R
KR
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
Lebedenko
E N
EN
Boni
I V
IV
Berlin
Y A
YA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D004742:Enhancer Elements, Genetic; D004926:Escherichia coli; D015870:Gene Expression; D005796:Genes; D005813:Genes, Synthetic; D005822:Genetic Vectors; D015186:Hominidae / Q000235:genetics*; D006801:Humans; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1 / Q000096:biosynthesis* / Q000235:genetics; D007438:Introns; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010442:Peptide Chain Initiation, Translational; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods; D011387:Prokaryotic Cells; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000737:chemistry; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D012795:Sialoglycoproteins / Q000096:biosynthesis* / Q000235:genetics
D017931:DNA Primers; C507763:IL1RN protein, human; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1; D012333:RNA, Messenger; D012795:Sialoglycoproteins