Cloning and characterization of directly amplified antiviral gene interferon alpha-2b (HulFNalpha-2b) from human leukocytes chromosomal DNA.
Interferons are cytokines that confer resistance to viral infection and inhibit cellular proliferation. The interferon alpha gene from human blood samples was amplified, cloned and expressed in E. coli (BL21). Leukocyte chromosomal DNA was used as a source of template DNA. Using specific primers, the gene for HulFNalpha-2b was amplified and inserted into the E. coli vector, pET21b, by ligation of the Hindlll and BamHI linkers of the vector and insert. The insert was further analyzed by PCR, DNA restriction mapping and sequencing, and expressed in a suitable E. coli strain. The production of this important cellular protein in the laboratory has significant applications in production of the recombinant pharmaceutical proteins.
Behravan
Javad
J
Pharmaceutical Biotechnology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. j_behravan@hotmail.com
Ahmadpour
Hassan
H
D002875:Chromosomes / Q000378:metabolism*; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000096:biosynthesis* / Q000235:genetics*; D007962:Leukocytes / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins; D004247:DNA