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Method for preparing porcine reproductive and respiratory syndrome resistant transgenic pig

A technology of transgenic cells and PRRS virus, applied in biochemical equipment and methods, genetic engineering, hybrid cell preparation, etc., can solve problems such as immunosuppression, high lethality, and economic losses in the poultry industry

Inactive Publication Date: 2011-07-06
BEIJING GEFUCURE BIOTECHNOLOGY LIMITED COMPANY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Infectious bursal disease (IBD) can cause severe immunosuppression and high lethality in chicken flocks, resulting in huge economic losses to the poultry industry

Method used

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  • Method for preparing porcine reproductive and respiratory syndrome resistant transgenic pig
  • Method for preparing porcine reproductive and respiratory syndrome resistant transgenic pig
  • Method for preparing porcine reproductive and respiratory syndrome resistant transgenic pig

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0060] Embodiment 1, the construction of RNAi vector

[0061] 1. Screening of siRNA interference targets

[0062] 1.1 Selection of RNA interference target genes

[0063] In the past five years, more than 85% of the PRRS viruses prevalent in my country belong to the JXwn strain, which is a highly pathogenic strain. The present invention selects the main structural protein genes (ORF5, 6, 7) and RNA polymerase gene (ORF1b) of the virus as interference target genes.

[0064] 1.2 Design and synthesis of siRNA sequence

[0065] Although the JXwn strain type is the target, in order to maximize the utilization of the selected interference target and avoid limiting the RNAi effect due to sequence differences among different strains, the target sequence of siRNA should be selected as much as possible among different strains. Conserved area. The selection criteria should be completely consistent sequences among different strains. If this requirement cannot be met, at least one misma...

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Abstract

The invention provides a method for preparing a porcine reproductive and respiratory syndrome resistant transgenic pig. The method comprises the following steps of: preparing a transgenic cell of DNA (Deoxyribose Nucleic Acid) of shRNA (short hairpin Ribose Nucleic Acid) which contains a code and takes a porcine reproductive and respiratory syndrome virus such as ORF1b, ORF5, ORF6 or ORF7 as an interference target gene; obtaining a cloned embryo by taking the transgenic cell as a nuclear transplantation donor cell and an excised oocyte as a nuclear transplantation recipient cell through a nuclear transplantation technology; and transplanting the cloned embryo into the uterus of livestock with a non-operative method for performing gravidity to obtain the transgenic pig. The transgenic pig obtained by the method has remarkable porcine reproductive and respiratory syndrome resistance.

Description

technical field [0001] The invention relates to animal transgenic technology, in particular to a method for preparing transgenic pigs resistant to PRRS. Background technique [0002] Porcine Reproductive and Respiratory Syndrome (PRRS), commonly known as PRRS, is an infectious disease caused by Porcine Reproductive and Respiratory Syndrome Virus (PRRSV). The disease was first discovered in the United States in 1987. In 2006, highly pathogenic PRRS broke out in pig herds in some provinces and cities in my country. Compared with the classic PRRS virus, the gene of this virus has undergone a large mutation and has many Strong pathogenicity. [0003] The economic losses caused by PRRS in the PRRSV pandemic have long been known. It is estimated that PRRS costs US swine producers $560 million per year (Cho and Dee, 2006). my country is a big pig raising country, accounting for almost half of the world's total pig raising. With the development of pig intensification and large-sc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/02C12N15/11
CPCC12N2310/14A01K67/0275A01K2267/02C12N15/1131A01K2217/00A01K2227/108
Inventor 鲍永华郭永臣李秋燕汤波李宁
Owner BEIJING GEFUCURE BIOTECHNOLOGY LIMITED COMPANY
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