Efficiently-expressed porcine interleukin-2 gene and application of expression protein of porcine interleukin-2 gene

A high-efficiency expression technology of interleukin, which is applied in the field of highly expressed porcine interleukin-2 gene and its protein expression, can solve the problems of unsatisfactory immune effect of inactivated vaccines, low expression level and activity of pIL-2, etc. Achieve the effect of enhancing humoral immunity and cellular immune response and reducing economic losses

Inactive Publication Date: 2013-07-10
INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the problems of unsatisfactory immune effect of porcine reproductive and respiratory syndrome inactivated vaccine and low expression level and activity of pIL-2 after porcine interleukin-2 gene transfection, the present invention provides a highly expressed porcine leukocyte interleukin-2 gene

Method used

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  • Efficiently-expressed porcine interleukin-2 gene and application of expression protein of porcine interleukin-2 gene
  • Efficiently-expressed porcine interleukin-2 gene and application of expression protein of porcine interleukin-2 gene
  • Efficiently-expressed porcine interleukin-2 gene and application of expression protein of porcine interleukin-2 gene

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Experimental program
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Embodiment Construction

[0045] 1.1 Artificial synthesis of inserted genes

[0046] Refer to the Rabbit β-Globin Intron II gene sequence published by GenBank (GenBank No: V00882), and introduce it upstream Sst I enzyme cutting site and a sequence of pCMV immediate early promoter, introduce T7 promoter sequence and Eco The RI restriction site, this gene sequence was spliced ​​together and handed over to TaKaRa Company for artificial synthesis. The specific coded nucleotide sequence containing the Rabbit β-Globin Intron II gene is shown in sequence 1 in the sequence table, and cloned into the T vector.

[0047] A segment of the pCMV immediate early promoter sequence is 5'-GTTTAGTGAACCGTCAGATCGCCTGGAGACGCCATCCACGCTGTTTTGACCTCCATAGAAGACACCGGGACCGATCCAGCCTCCGC-3',

[0048] The T7 promoter sequence is 5'-AATACGACTCACTATAGGG-3';

[0049] 1.2 pMVAX1 ? construction and identification of

[0050] from the T carrier Sst I / Eco The target gene was cut with RI double enzymes and recovered from the gel, a...

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Abstract

The invention relates to the field of gene expression and in particular relates to an efficiently-expressed porcine interleukin-2 gene. The nucleotide sequence of the porcine interleukin-2 gene is expressed as a sequence 2; the gene of the sequence 2 is cloned to a recombinant plasmid acquired by an eukaryotic expression vector; the expression protein of the porcine interleukin-2 gene plays a role in obviously enhancing the immunity of an inactivated vaccine of porcine reproductive and respiratory syndrome; a pIL-2 protein gene is expressed by an eukaryotic plasmid pMVAX<0>-pIL-2 containing the sequence 2 in a sequence table, so that the limitations of low expression quantity and poor activity in the process of expressing the pIL-2 via a pMVAX<0> vector are broken, and the expression quantity is improved by 5.7 folds; and pig experiments prove that the expression activity of the expression protein pIL-2 of the gene in the sequence 2 in the sequence table is high, and the response of humoral immunity and cellular immunity of a pathogenicity PRRSV (Porcine Reproductive and Respiratory Syndrome Virus) inactivated vaccine can be obviously improved, so that the expression protein pIL-2 is specially used for preventing diseases caused by high pathogenicity PRRSV and reducing financial loss of pig farms.

Description

technical field [0001] The invention relates to the field of gene expression in biotechnology, in particular to a high-efficiency expressed porcine interleukin-2 gene, and also relates to the application of the expressed protein. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS), commonly known as "pig blue ear disease", is caused by porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome virus, PRRSV). Pig abortion, return to oestrus, stillbirth or weak piglets, piglet death and different degrees of respiratory symptoms in pigs of various ages can destroy the immune system of pigs and cause mixed infections or secondary infections. The disease broke out in my country at the end of 1995 and spread rapidly to many provinces across the country. In many large-scale pig farms, the positive rate of PRRS is very high, and some can reach more than 80%. Since 2006, highly pathogenic PRRSV caused by PRRSV m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/26C12N15/85C07K14/55A61K39/39
Inventor 杜以军齐静王金宝吴家强黄保华孙文博丛晓燕郭立辉任素芳陈蕾李俊时建立吕伟
Owner INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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