Molecular cloning of human interleukin 2 cDNA and its expression inE. coli

Abstract

polypeptide had a molecular weight of 15,000 daltons and accounted for more than 10% of the total E. coli proteins in fully induced cells; it was biologically active in the T-cell specific DNA synthesis assay, even after recovery from a SDS-containing polyacrylamide gel.

Isolation of a functional human interleukin 2 gene from a cosmid library by recombination in vivo

“…4B). It is also in agreement with restriction maps derived from published sequences of part of this region, except for one Xbal site , Degrave et al, 1983, Holbrook et al, 1984. There are no discrepancies between fragments of human placental DNA digested with EcoRI, Hindlll, Bglll and Pvull hybridizing to IL2 cDNA (not shown) and the fragments contained in pHIL49.…”

“…The coding sequence of the human IL2 gene has been cloned from this cell line and from lymphoid tissues , Maeda et al, 1983. Recently the isolation of chromosomal DNA fragments containing the IL2 sequences by conventional screening of phage A. libraries has been described Degrave et al, 1983;Holbrook et al, 1984 ). To study expression and modification of IL2 in homologaus and heterologous animal ceJls in detail we decided to isolate the region containing the chromosomal gene and flanking sequences from a human cosmid library.…”

Enhancement of translational efficiency by the Escherichia coli atpE translational initiation region: its fusion with two human genes

“…The segment of 30 bp or less upstream from the atpE SD plays a primary role in determining this high Ievel oftranslation, just as was shown previously for the atpE gene itself (McCarthy et al, 1985). The maximal levels of synthesis of the two human genes in E. coli obtained in the present work using the atpE sequence are at least as high as those previously reported by others (Devos et al, 1983~ Remaut et al, 1983.…”

“…This observation, taken tagether with the competitive inhibitory effect of expression of the cloned foreign genes upon the expression of other cellular genes (Fig. 2), and also the Ievel of expression attained (Devos et al, 1983;Remaut et al, 1983 ), indicates that IL2 and IFNß were both expressed at very near the physiological Iimits of overproduction for these two genes in E. coli.…”

The purification of eukaryotic polypeptides synthesized in Escherichia coli

“…Folded eukaryotic products with correct disulphide bonds can therefore be produced, as demonstrated for proinsulin (Emerick et al, 1985) and EGF (Oka et al, 1985). Periplasmic secretion It was suggested that E. coli cannot process eukaryotic signal peptides efficiently (Devos et al, 1983) but more recent evidence is contradictory. The eukaryotic signal of IgG light chain both initiated secretion and was correctly processed (Zemel-Dreasen & Zamir, 1984) but when the prokaryotic ,J-lactamase signal sequence was placed in front of the IgG light chain signal sequence only the bacterial signal peptide was processed during secretion.…”