Fishy taste susceptible gene of quail egg and application of fishy taste susceptible gene as molecular marker

A molecular marker, fishy smell technology, applied in application, genetic engineering, plant genetic improvement and other directions, to achieve the effect of improving economic efficiency, simple operation and reducing expenditure

Inactive Publication Date: 2013-01-30
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are no reports on the key genes that affect

Method used

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  • Fishy taste susceptible gene of quail egg and application of fishy taste susceptible gene as molecular marker
  • Fishy taste susceptible gene of quail egg and application of fishy taste susceptible gene as molecular marker
  • Fishy taste susceptible gene of quail egg and application of fishy taste susceptible gene as molecular marker

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Experimental program
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Embodiment 1

[0028] Molecular cloning of embodiment 1 quail FMO3 gene

[0029] 1. Preparation of total RNA from quail liver tissue

[0030] Total RNA was extracted using Trizol one-step method

[0031] (1) Preparation: Prepare Trizol, chloroform, isopropanol, 75% ethanol and RNA solution DEPC water.

[0032] Add 1.6ml Trizol to a 2ml imported centrifuge tube in advance and store at 4°C for 1-2 hours.

[0033] (2) Tissue homogenate: Grind the tissue sample with a mortar until it becomes powdery, and then transfer it to a centrifuge tube filled with Trizol. Shake to dissolve the tissue sample in Trizol as much as possible until the solution is clear. Place at room temperature 15-30°C for more than 10 minutes to increase RNA yield.

[0034] (3) Phase separation: Add 0.32ml of chloroform to the centrifuge tube containing the lysate, and vibrate the centrifuge tube vigorously for 15 seconds. Centrifuge at 12000rmp for 15 minutes at 4°C.

[0035] (4) Precipitation: Transfer the supernatant...

Embodiment 2

[0127] The sequencing results were analyzed with Chromas2.23 and DNAMAN5.2.2. According to the sequencing results, the sequences were spliced. Example 2 Establishment of DNA sequencing detection method and determination of polymorphic sites

[0128] 1. PCR amplification

[0129] Select Shendan’s self-separated male and female eggs and use two pure-line quails of male and female quail as experimental materials, take 20 μL quail anticoagulant whole blood, use traditional phenolic extraction method to extract blood total DNA, and use 1.5% agarose gel DNA quality was detected by electrophoresis.

[0130] Primers were designed according to the cDNA sequence of the quail FMO3 gene, the sequence is as follows:

[0131] F primer (upstream primer): 5'-GTGAAGGAATTCAGAGAAACA-3'

[0132] R primer (downstream primer): 5'-CTGAAACACCTTGACTGCC-3'

[0133] Use quail genomic DNA as a template, use primers F primer and R primer for PCR amplification, and use a 15 μL reaction system:

[013...

Embodiment 3

[0144] Example 3 Detection of the distribution of the Q319X site C / T mutation in paternal and maternal populations

[0145] According to the method of Example 1, the distribution of the C / T mutation at the Q319X site of the FMO3 gene was detected in two pure line populations of the male and female quail mating lines of the Shendan self-separating eggs, and the detection results are shown in Table 1.

[0146] Table 1 Distribution of C / T mutation at Q319X site of FMO3 gene in paternal and maternal populations

[0147]

[0148] According to the results in Table 1, it can be seen that the fishy smell sensitive gene T and the sensitive genotype TT individuals were detected in both the male and female quail mating lines of Shendan self-separating eggs, indicating that the two groups of quail were fed with 4000mg / Kg A small number of sensitive genotype individuals will produce fishy-smelling eggs with high trimethylamine content in the diet of the experimental group, and molecular...

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Abstract

The invention relates to a fishy taste susceptible gene of a quail egg and application of the fishy taste susceptible gene as a molecular marker. A nucleotide sequence of the fishy taste susceptible gene FMO3 (Flavin Containing Monooxygenase3) is represented by SEQ ID NO: 1. A Q319X mutational site is served as an SNP (Single Nucleotide Polymorphism) molecular marker for detecting the fishy taste susceptible gene of the quail egg. The fishy taste susceptible gene of the quail egg, provided by the invention, is an auxiliary molecular marker of the quail egg with a fishy taste character, so that a molecular genetic marker selecting method with high efficiency is provided for accelerating the improvement of the flavor of the quail egg. The method greatly reduces the molecular breeding expenditure, effectively gets rid of the problem that the quail egg has fish taste in the production, enlarges the range of the selective feed raw materials of the quail, reduces the feeding cost, and improves the economic benefit of the quail breeding. The method is simple to operate, cheap in cost, short in detection time, high in accuracy, and capable of realizing automatic detection, thereby playing a great role the in quail breeding.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a quail egg fishy smell sensitive gene and its application as a molecular marker. Background technique [0002] Over the past 30 years of reform and opening up, the animal husbandry industry has achieved rapid development relying on the support and encouragement policies of the state, providing the people with a variety of rich livestock and poultry products. With the gradual improvement of material life, the people are no longer satisfied with having enough to eat, and put forward higher requirements for various livestock and poultry products in terms of product quality. The total number of quails raised in my country is 200 million, of which egg-type quails account for three-quarters. Quail eggs have been on people's tables as a common poultry egg product. [0003] Quail eggs have high nutritional value, are rich in various essential nutrients for human body, have excellen...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N15/11C12Q1/68C12N9/02
Inventor 杨宁郑江霞墨锋涛
Owner CHINA AGRI UNIV
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