Recombinant expression sheep interferon-tau BB8 gene and preparation method thereof

Abstract

The invention provides a recombinant expression sheep interferon-tau BB8 gene and a preparation method thereof. The nucleotide sequence of the recombinant expression sheep interferon-tau BB8 gene is as shown in SEQ ID NO.1, DNA molecules of the recombinant expression sheep interferon-tau BB8 gene are optimized and replaced with codons preferred by escherichia coli, that is, the recombinant expression sheep interferon-tau BB8 gene is obtained by inserting an NdeI site at the 5' end of a natural sheep interferon-tau BB8 gene sequence and inserting a termination codon and a HindIII site at the 3' end of the natural sheep interferon-tau BB8 gene sequence. The active sheep interferon-tau BB8 protein is expressed in an escherichia coli system for the first time, escherichia coli preferred codons are selected for the artificially designed and synthesized sheep interferon-tau BB8 gene, identification and expression of the codons in the escherichia coli are facilitated, and therefore, the purpose of efficient expression can be achieved. The expression system disclosed by the invention is beneficial to correct folding of the recombinant protein and promotes expression of the sheep interferon-tau BB8 in a soluble form, and the protein is subjected to one-step purification under a non-denaturation condition to obtain a high-purity product.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, in particular to a recombinantly expressed goat interferon-τBB8 (Goatinterferon tau BB8, GoIFN-τBB8) gene and a preparation method thereof. Background technique [0002] Interferon-τ (Interferon-tau, IFN-τ) is an acidic, low-molecular-weight protein secreted by ruminant embryo trophoblast cells. It was originally called embryo trophoblast protein-I. plays an important role in the process [1,2] . Although IFN-τ is not induced by viral infection, IFN-τ is also classified as a type I interferon based on its amino acid sequence homology with IFN-α and IFN-ω [3] , and has similar anti-proliferation and anti-viral activities as the latter two, and when used for disease treatment, compared with the former two, it has almost no toxic side effects [4 -6]. Bazer et al. reported that ovine interferon-τ had the same effect as human interferon-α on preventing the replication of human and felin...

AI Technical Summary

Problems solved by technology

Partially soluble expression of sheep IFN-τ was achieved by fusion expression, and the activity after purification was 1×10 6 IU / mg, less active

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