Novel process for preparing recombinant chicken interferon alpha with strong activity and high expression quantity
An alpha interferon, high expression technology, applied in the field of interferon, can solve problems such as unfavorable purification, low protein expression, and achieve the effect of improving protein expression and high expression
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[0087] A new process for preparing recombinant chicken interferon alpha with strong activity and high expression, comprising the following steps:
[0088] S1. Construction of a cloning vector for recombinant chicken interferon-α
[0089] S11. Extract RNA from chicken lymphocytes, reverse transcribe the RNA to obtain chicken α-interferon cDNA, use ChIFNα primers to perform PCR amplification on cDNA, obtain chicken α-interferon gene PCR product, connect with plasmid pJET, and transform , to obtain the recombinant plasmid pJET-ChIFNα, which specifically includes the following steps:
[0090] S111. Extract RNA from chicken lymphocytes: aseptically collect 5 mL of chicken whole blood intravenously, add 5 mL of 1% sterile sodium heparin 1640 culture solution to obtain whole blood containing heparin sodium, and pre- Add 5 mL of lymphocyte separation medium, slowly add 5 mL of whole blood containing heparin sodium on top of the separation liquid, centrifuge at 1500 rpm for 20 minutes a...
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