Synthesis, Cloning And Expression In E. Coli Of Mouse Interleukin 4 Gene

The total DNA synthesis, cloning and expression in E. coli of mouse interleukin 4 (MIL —4) gene is described.The MIL — 4 gene, consists of 442 base-pairs (bp), was devided into 29 oligonucleotide fragments and synthesized on ABI 381A DNA synthesizer. It is covenient for the gene cloning with Hind Ⅲ , EcoRI sites and an initial codon (ATG) at the 5' —terminus, and two terminal codons (TAA, TAG) and Hind Ⅲ cohesive end at the 3' —terminus of the synthetic gene. The synthetic oligonucleotides were assembled in two groups and inserted into plasmid pUC12 respectively, resulting in two recombinant plasmids, pUC12A and pUC12B. The pUC12A plasmid consists of the first half gene fragment of MIL—4. The pUC12B plasmid consists of the second half gene fragment of MIL —4. The final identification of the two gene fragments was by DNA sequencing. Ligation of the first and last half gene fragments of MIL—4, we ob-...