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Primer combination for identifying creeping character of chicken and application thereof

A technique for combining primers and identifying chickens, which is applied in the field of molecular biology, can solve problems such as low degree of automation, long analysis and determination cycle, complicated sampling operation, etc., achieve wide application range, improve hatchability and uniformity of growth and development, Simple operation effect

Active Publication Date: 2016-06-15
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to conventional breeding methods, the selection of creeping traits is mainly determined by the length of the shank, which takes a long period and costs a lot
In addition, conventional morphological and cytological methods for detecting Cp homozygous lethal embryos and normal dead embryos have complex sampling operations, long analysis and measurement cycles, low automation, and insufficient detection accuracy.

Method used

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  • Primer combination for identifying creeping character of chicken and application thereof
  • Primer combination for identifying creeping character of chicken and application thereof
  • Primer combination for identifying creeping character of chicken and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Design and screening of embodiment 1 primer combination

[0029] The design method of the primer combination, such as figure 1 shown.

[0030] 1.1 Primer design

[0031] The creeping genes used to design primers have been identified through high-throughput sequencing and bioinformatics analysis, which have important biological significance and potential application value. The gene sequence was also obtained through bioinformatics analysis. Due to the large missing sequence and multiple PCR reactions, in addition to the specificity, sensitivity, and amplification efficiency of the primers, the position of the primer design (the middle sequence of the missing fragment and the upstream and downstream sequences of the missing sequence), two PCR product size, Tm consistency of two pairs of primers, primer ratio, typing efficiency, etc. One multiplex PCR reaction can accurately distinguish individuals of different genotypes. This combination of primers simplifies the ope...

Embodiment 2

[0057] The establishment of embodiment 2 multiplex PCR method and Cp genotyping

[0058] 1. Analysis of Cp genotypes in different individuals

[0059] 1. Experimental materials. The experimental flock construction and sample collection were completed in the China Agricultural University Poultry Genetic Resources and Breeding Experimental Base. Xingyi bantam chicken is a valuable poultry genetic resource, with special prostrate traits, which are controlled by a single dominant gene Cp, which is a dominant homozygous lethal gene on autosomal, gene heterozygosity leads to prostrate traits, embryonic death was concentrated on the fourth day after hatching. In this experiment, the progeny of prostrate roosters and prostrate hens were selected as the experimental subjects.

[0060] 2. Genomic DNA extraction

[0061]Whole chicken embryos hatched for 4 days were collected and genomic DNA was extracted with a tissue extraction kit. Blood was collected from wing veins of chickens a...

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Abstract

The invention relates to the field of molecular biology, in particular to a primer combination for identifying the creeping character of chicken and the application thereof.The primer combination comprises primer sequences as shown in the SEQ ID No.1, SEQ ID No.2, SEQ ID No.3 and SEQ ID No.4.The creeping character of chicken can be identified by means of the primer combination.Specifically, the genomic DNA of an individual to be tested is taken as the template, and analysis is conducted with the multiple PCR method by means of the primer combination; if one product is obtained through multiple PCR amplification and the sequence size of the product is 224 bp, the individual to be tested is a creep gene dominant homozygote (Cp / Cp), and the individual is of the lethal type; if two products are obtained through multiple PCR amplification and the sequence sizes of the products are 438 bp and 224 bp respectively, the individual to be tested is a creep gene heterozygote (Cp / +), and the individual has the creeping character; if only one product is obtained through multiple PCR amplification and the sequence size of the product is 438 bp, the individual to be tested is a recessive homozygote (+ / +), and the individual is of the wild type.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a combination of primers for identifying chicken prostrate traits and an application thereof. Background technique [0002] So far, eight dwarf genes have been found in chickens, which are respectively located on sex chromosomes and autosomes. Among them, the sex-linked dwarf gene dw has been studied more, and the gene has been precisely located and its mechanism of action has been clarified ( Agarwal, S.K., L.A. Cogburn, and J. Burnside. 1994. Dysfunctional growth hormone receptor ina strain of sex-linked warf chicken: evidence for a mutation in the intracellular domain. J. Endocrinol. 142:427-434.). There are many dwarf genes on autosomes, among which Cp gene (Cp: Creeper, creeping type) is an important dominant homozygous lethal gene on autosomes (Landauer, W., and L.C.Dunn.1930.Studies on the Creeperfowl .I.Genetics.J.Genet.23:397-413.), the gene controls the prostrate trait...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2600/124C12Q2600/158C12Q2600/16C12Q2537/143
Inventor 侯卓成金四华杨宁郑江霞李俊英
Owner CHINA AGRI UNIV
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