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Mitochondria molecular hereditary feature mark and method for identifying pure boer goat

A Boer goat, genetic marker technology, applied in sugar derivatives, organic chemistry, fermentation, etc., can solve the problems of high price, inaccuracy, and high price of purebred

Inactive Publication Date: 2005-01-19
SHANGHAI GENON BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the original breeds of Boer goats were imported from abroad in the early stage, the relative price was relatively expensive, which led to relatively high prices for the original breeds and pure breeds that were brought back to China for breeding and breeding.
At present, the traditional morphological identification method is used to identify purebred Boer goats, which is very inaccurate. As a result, criminals use the first-generation and second-generation Bourbon hybrid sheep to pretend to be Boer goats to deceive customers.

Method used

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  • Mitochondria molecular hereditary feature mark and method for identifying pure boer goat
  • Mitochondria molecular hereditary feature mark and method for identifying pure boer goat
  • Mitochondria molecular hereditary feature mark and method for identifying pure boer goat

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Goat Cell Mitochondrial DNA Preparation

[0063] Goat mitochondrial DNA can be prepared separately by the following three methods:

[0064] method 1.

[0065] Take a small amount of goat ear tissue or blood. Total DNA (mitochondrial DNA is included in the total DNA) was extracted from the ear tissue according to conventional methods, and dissolved with TE.

[0066] Method 2.

[0067] Add 3-10 times the volume of distilled water to the blood, treat it in boiling water for 10 minutes, centrifuge and take the supernatant as a template.

[0068] Method 3:

[0069] Animal mitochondria are first purified, and then mitochondrial DNA is extracted and purified by conventional genomic DNA extraction methods.

[0070] The DNA prepared by the above three methods can be used in the amplification reaction. Among them, method 2 is the most convenient, but the quality of template DNA is slightly worse. Method 3 is the most cumbersome, but the template DNA is the purest.

Embodiment 2

[0072] PCR Amplification of Mitochondrial D-loop Region in Goat Cells

[0073]According to Genebank goat cytochrome b gene (Capra hircus mito...[gi:17298008]) and 12srRNA gene (Capra hircus Phe-...[gi:337103]) respectively synthesize the following 3 pairs of PCR primers for three rounds of nested PCR Amplify.

[0074] Sequence (5'-3')

[0075] 1. Goatmtb528: 5' CCG ATT CTT CGC CTT CCA C 3' (SEQ ID NO: 3)

[0076] Goatmt12s783: 5'GC CCA TTT CTT CCC ATT CCA3' (SEQ ID NO: 4)

[0077] 2. Goatmtb570: 5' AGC CCT CGC CAT AGT CCA CCT 3' (SEQ ID NO: 5)

[0078] Goatmt12s699: 5'GGT TTG CTG AAG ATG GCG GTA 3' (SEQ ID NO: 6)

[0079] 3. Goatmtb1020: 5'ACA GCC AGT CGA ACA TCC CTA C 3' (SEQ ID NO: 7)

[0080] Goatmt12s142: 5'TAC TCA CCG GGG CGT GGA TG3' (SEQ ID NO: 8)

[0081] Note: b528 in the primer name indicates that it corresponds to the 528th position of the cytochrome b gene, 12s783 indicates that it corresponds to the 783rd position of the 12s rRNA gene, and so on.

[0082] T...

Embodiment 3

[0112] Restriction Enzyme Map of PCR Products in D-loop Region of Mitochondrial in Goat Cells

[0113] The Taq I restriction endonuclease from TaKaRa Company was used, and the reaction conditions recommended by the manufacturer were carried out. The total reaction volume was 20 μl. The PCR product of the third round was directly digested for 2-3 hours, and then bromophenol blue loading buffer was added to terminate the reaction. And electrophoresis in 2% agarose gel.

[0114] Results and Analysis

[0115] The 1614bp amplified product (SEQ ID NO: 1) of purebred Boer sheep contains an additional specific enzyme cutting site at position 911 ( Figure 2A ), so the resulting lengths were 10, 170, 533, and 901 after enzyme digestion.

[0116] The 1614bp amplified product (SEQ ID NO:2) of other goats only contains TaqI site ( Figure 2B ), so the resulting lengths were 10, 533, and 1071 after enzyme digestion. Therefore, purebred Boer sheep can be identified conveniently, quickly...

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Abstract

The invention discloses a whole-bred Pole goat mitochondria DNA nucleotide sequence in ring D domain, and oligonucleotide primer designed on the basis of the sequence, the invention also provides the process for judging the whole-bred Pole goats.

Description

technical field [0001] The invention belongs to the technical field of detecting biological species by means of molecular biology methods. More specifically, it relates to a genetic marker for mitochondrial DNA molecular characteristics of Boer goats, and a method for detecting purebred Boer goats based on the markers. Background technique [0002] In the production structure of meat products, beef and mutton are low in fat and high in protein. They are not only high-quality meat products, but also the main raw materials for halal meat products. [0003] Boer goat (Capra hircus, Boer) is recognized as an excellent meat breed in the world. It has excellent characteristics such as large individual, fast initial growth rate, wide adaptability, good reproductive performance, and high meat yield. China is a big agricultural country, and animal husbandry occupies an extremely important position in agriculture. However, there is a considerable gap compared with developed countrie...

Claims

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Application Information

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IPC IPC(8): C07H21/00C12P19/34C12Q1/68
Inventor 成国祥赵建阳彭进蔡引凤王述宇张锁林
Owner SHANGHAI GENON BIOENG
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