Composite porcine alpha-IFN gene and recombinant vector thereof

Abstract

The invention relates to a composite porcine alpha-IFN gene and a recombinant vector thereof, and belongs to the technical field of genetic engineering biological products. The newly designed gene sequences of composite porcine alpha-IFN are recombined to pPICZ alpha-A vectors, and the vectors are electronically transformed into yeast competent cells; and the positive identification, and the screening and the induced expression of high copy yeast strains are carried out. A pichia pastoris expression system is used for expressing foreign genes so as to be favorable for the commercial production of porcine IFN. Compared with the natural IFN which is expressed by yeast, the expression level of the protein which is expressed by genome increases by 3.17 times. The fluorescent quantitative PCR result shows that the level of PRVmRNA after PK-15 cells treated by 10-6mg / ml of the composite porcine alpha-IFN are re-infected with PRV is obviously lower than the level of PRVmRNA after the PK-15 cells which are not treated by the composite porcine alpha-IFN are infected with the PRV, and the level of PRVmRNA decreases by 37 times.

Description

1. Technical field [0001] The invention discloses a gene sequence of a compound porcine alpha interferon and its recombinant carrier, belonging to the technical field of genetic engineering biological products. Compared with natural porcine interferon alpha, it has stronger antiviral activity and can be used for the prevention and early treatment of various viral diseases in pigs. 2. Background technology [0002] Interferon is one of the most important cytokines in the animal body. It has broad-spectrum and high-efficiency anti-virus functions and plays a key role in regulating the immune system. As early as 1986, Lefevre et al. had done detailed research on porcine alpha interferon gene and porcine alpha interferon. A fragment containing α-interferon gene was obtained. Sequence analysis showed that the fragment had a continuous open reading frame containing 190 codons, a 5' region of about 100 nucleotides upstream of the initiation codon, and a TATTTAA sequence ( It has ...

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Problems solved by technology

[0005] The production of porcine IFN-α induced by inducers is expensive due to many limitations such as few sources, high cost, and complicated purification process, which greatly limits clinical and scientific applications.

In the prokaryotic expression system, its products mainly exist in the form of inclusion bodies without biological activity. If you want to obtain recombinant proteins with biological activity, you must carry out tedious follow-up processing such as denaturation and renaturation of the inclusion bodies, and the recombinant protein in the follow-up processing work The amount of loss is large and the technical level of the staff is also very high, which is not conducive to industrial production

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