Unlock instant, AI-driven research and patent intelligence for your innovation.

DOCK2 gene fragment used as molecular marker for detecting immune character of pig and application of DOCK2 gene fragment

A molecular marker and trait technology, applied in DNA/RNA fragmentation, recombinant DNA technology, microbial determination/inspection, etc., can solve the problems of reduced slaughter rate, low disease control ability, and pig death, etc., and achieves fast and low cost. , detect accurate effect

Active Publication Date: 2015-11-04
HUAZHONG AGRI UNIV
View PDF0 Cites 13 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the development of the pig industry is also facing many challenges, among which the prominent problem is that the ability to control certain diseases is still low, and there are still many problems in the links of disease monitoring, diagnosis, prevention and extermination. Therefore, in the breeding production process The disease problem is one of the key issues affecting the development of the pig industry
The outbreak of diseases in the process of pig breeding can directly lead to the death of pigs and reduce the slaughter rate, which in turn affects the economic benefits of pig farming
At the same time, the extensive use of drugs to prevent diseases increases the cost of farming on the one hand, and on the other hand, it also leads to the emergence of drug resistance of bacteria, making it more difficult to prevent and control diseases in farms. The extensive use of drugs also makes pork food safety problems prominent.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • DOCK2 gene fragment used as molecular marker for detecting immune character of pig and application of DOCK2 gene fragment
  • DOCK2 gene fragment used as molecular marker for detecting immune character of pig and application of DOCK2 gene fragment
  • DOCK2 gene fragment used as molecular marker for detecting immune character of pig and application of DOCK2 gene fragment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Cloning of Example 1 Intron Sequence of Pig DOCK2 Gene

[0031] (1) Extraction of total DNA from ear tissue of Duroc×Erhualian F2 population by phenol extraction

[0032] 1) Grind the ear tissue of Duroc × Erhualian F2 population (provided by Guangdong Huanong Wens Animal Husbandry Co., Ltd.) in liquid nitrogen, add an equal volume of 1×SET (1mL), proteinase K (10ng / mL) To a final concentration of 200ug / mL, sodium dodecyl sulfate (ie SDS, 10%) to a final concentration of 0.5%, shake well. Incubate overnight in a 55°C water bath for digestion.

[0033] 2) Add an equal volume of Tris-saturated phenol to the digested tissue sample, slowly invert the centrifuge tube for 15 minutes, centrifuge in a low-temperature refrigerated centrifuge at 4°C and 11,000 rpm for 10 minutes, and carefully transfer the supernatant to another centrifuge tube. on the corresponding mark.

[0034] 3) Add an equal volume of phenol / chloroform / isoamyl alcohol (volume ratio 25:24:1), slowly invert...

Embodiment 2

[0049] Embodiment 2: The establishment of CRS-PCR-RFLP genotype detection method

[0050] (1) Primer design

[0051] A pair of primers were designed for the polymorphic site A35-G35 to introduce a restriction site for genotype detection. The DNA sequence of the primer pair is as follows:

[0052] Forward primer: 5'-AGATAAGCCACCACGGAATGGAGAAATTGCgTAT-3' (the original base T at the 31st position of the primer sequence is mismatched into g, see the sequence table SEQ ID NO: 2)

[0053] Reverse primer: 5'-ATCCCTGGTCTGTGGCATGTCC-3' (see sequence listing SEQ ID NO: 3)

[0054] (2) PCR amplification

[0055] The total volume of the PCR reaction system is 20ul: double distilled water 10.5ul, 10x PCR Buffer 2ul, dNTP Mix 1.6ul, forward primer 0.4ul, reverse primer 0.4ul, rTaq enzyme 0.1ul, Duroc×Erhualian F2 population genomic DNA 5ul . Amplification conditions: pre-denaturation at 95°C for 1 min, denaturation at 95°C for 30 s, annealing at 59°C for 30 s, extension at 72°C for 15 s...

Embodiment 3

[0058] Example 3: Application of DOCK2 molecular marker typing method in association analysis of immune traits

[0059] (1) Correlation analysis between DOCK2 molecular marker typing results and immune traits

[0060] The original source of the experimental pigs used for the detection and analysis of the association between genotype and immune traits was the F2 population (conventional breed) of the Duroc × Erhualian hybrid bred by Guangdong Huanong Wenshi Animal Husbandry Co., Ltd. The DNA used for genotyping was extracted from the ear samples of the F2 generation of the Duroc × Erhualian cross (the following text and the "F2 generation of the Duroc × Erhualian cross" in Table 1-8 are referred to as "pigs") for blood The blood for routine detection and flow cytometry analysis was collected from pigs aged 20, 33, 35 and 80 days. Pigs aged 20 and 33 were not inoculated with polymyocytes (PolyI:C), and piglets at 35 days were inoculated with polymyocytes Blood samples were coll...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the technical field of livestock molecular marker preparation and particularly relates to a DOCK2 gene fragment used as a molecular marker for detecting immune character of pig and application of the DOCK2 gene fragment. The molecular marker is obtained by cloning a DOCK2 gene of the pig; the molecular marker has a nucleotide sequence as shown in a sequence table SEQ ID NO:1; the basic group at the 35th site of the sequence is provided with an A35-G35 basic group mutation; the A35-G35 forms BstZ17I-RFLP polymorphism because the mutation is introduced to the fourth basic group position backwards counted from a forward primer. The invention provides a novel SNP molecular marker for assisted selection of marking of the immune character of the pig.

Description

technical field [0001] The discovery belongs to the technical field of livestock molecular marker preparation, and specifically relates to DOCK2 gene fragments as molecular markers for pig immune traits and their application. Background technique [0002] my country has a long history of pig raising, rich in pig raising experience and variety resources, and the number of pigs raised ranks first in the world, and has promoted the development of the world's pig raising industry. However, the development of the pig industry is also facing many challenges, among which the prominent problem is that the ability to control certain diseases is still low, and there are still many problems in the links of disease monitoring, diagnosis, prevention and extermination. Therefore, in the breeding production process The disease problem in pigs is one of the key issues affecting the development of pig industry. The outbreak of diseases in the process of pig breeding can directly lead to the...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 李新云倪娟李世军赵书红曹建华刘向东谢胜松
Owner HUAZHONG AGRI UNIV