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Home >> Biotechnology and Genomics >> Isolation, Sequencing and Synthesis of Genes >> Total Synthesis of a Human Leukocyte Interferon Gene

Total Synthesis of a Human Leukocyte Interferon Gene
Interferon is a protein, which has a property of inhibiting viral infection and cell proliferation and thus can be used for treatment of viral infection and malignancies (cancer). These properties have 'generated great interest in human interferons (IFNs). At least three kinds of interferon genes are known in human genome: (i) leukocyte interferon genes. (IFN-α genes), (ii) fibroblast interferon genes (IFN-βgenes), and (iii) immune interferon gene (IFN- γ genes). Genes of different kinds of interferons differ and details of base sequences are now known for all of them.

In 198O, Weismann and coworkers published the nucleotide sequence of the first IFN-a gene derived from cloned IFN-a eDNA. On the basis of this information, total synthesis of this human leukocyte interferon gene, 514 base pairs long, was achieved and published in 1981. Since there are 514 base pairs in the gene, it involved synthesis of DNA strands with 1028 nucleotides in a predetermined way. This was the longest gene synthesized till 1982. The synthesis involved coupling of initial nucleotide onto a polyacrylamide resin to which further nucleotides could be added in pairs. Sixty six (66) oligonucleotide segments varying in size from 14 to 21 bases were synthesized, which were then arranged in a predetermined way and joined.

This gene was incorporated into a plasmid and was capable of synthesizing α-interferon. The 514 base pairs coded for 166 amino acids = 498 base pairs, and included the initiation and termination signals. There are no intron sequences present in this gene (introns are noncoding sequences, common in eukaryotic interrupted genes, that are known as split genes).

The plasmids with this newly synthesized gene were transferred into bacteria (E. coli), which are being utilized now for synthesis of interferon in industry. This has greatly reduced the cost of production of this drug (interferon) which was earlier sold at the .rate of Rs. 16 million per 50 mg.

 

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