ABSTRACT
Expression E. coli plasmid were constructed in which the human interleukin-4 (hIL4) synthetic gene is controlled by tac promoter. The expression level of the gene depends on the distance between RBS and the initial codon ATG, with the maximal production in case of the nine base pair distance. The recombinant protein, accumulated in the inclusion bodies, was solubilized, renaturated, and purified to homogeneous, biologically active preparation, the yield being 2 mg/g wet cells.
Subject(s)
Escherichia coli/genetics , Gene Expression , Genes, Synthetic , Interleukin-4/genetics , Base Sequence , Humans , Interleukin-4/isolation & purification , Interleukin-4/pharmacology , Molecular Sequence Data , Plasmids , Promoter Regions, Genetic , T-Lymphocytes/drug effectsABSTRACT
A modified H-phosphonate method was used to synthesize 32 oligodeoxyribonucleotides ranging in length from 23 to 28, which were enzymatically joined together to give the human interleukin 4 gene. The high degree of the oligonucleotide purity, achieved through the application of anion-exchange and reverse phase HPLC, ensures the high percentage of the desired sequence (about 75%) in the cloned DNA.