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The primers specific to cervus elaphus, c. nippon, c. canadensis and rangifer tarandus gene and the method to identify cervi parvum cornu species

An identification method, a technology of sika deer, applied in biochemical equipment and methods, microbe determination/inspection, DNA/RNA fragments, etc., can solve the problem of no introduction of velvet antler

Inactive Publication Date: 2007-03-21
KOREA INST OF ORIENTAL MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] As mentioned above, although there are many methods of using gene-specific primers to detect diseases or individuals, there has never been an example of introducing them into the selection of velvet varieties.

Method used

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  • The primers specific to cervus elaphus, c. nippon, c. canadensis and rangifer tarandus gene and the method to identify cervi parvum cornu species
  • The primers specific to cervus elaphus, c. nippon, c. canadensis and rangifer tarandus gene and the method to identify cervi parvum cornu species
  • The primers specific to cervus elaphus, c. nippon, c. canadensis and rangifer tarandus gene and the method to identify cervi parvum cornu species

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] (Example 1) Preparation of velvet antler sample

[0058] The blood and tissues of red deer, sika deer, red deer, and reindeer raised in Seoul Grand Park were used as the standard sample of antler. As other blood, the blood and blood of red deer, sika deer, and red deer raised on domestic individual farms and domestic The circulating dried velvet antler is used in experiments. Use QIAamp DNA microkit (QIAGEN Germany) to extract all the DNA isolated from the blood or tissue of deer antler according to the user's instructions.

Embodiment 2

[0059] (Example 2) Confirming the partial base sequence of the D-loop of velvet antler and determining the standard D-loop base sequence

[0060] In order to amplify the D-loop region of mitochondrial DNA (mtDNA) of red deer (Cervus elaphus), sika deer (Cervus nippon), red deer (Cervuselaphus canadensis) and reindeer (Rangiger tarandus) for base sequence analysis, three primers were used. The CST2 primers and CST39 primers described in sequence numbers 1 and 2 use the primers published in Polziehn et al. (1998), because the SeqR primer described in sequence number 3 cannot base the entire D-loop in one experiment. Sequence analysis is based on the result of base sequence analysis with CST39 primers for re-synthesis, and then completes the remaining base sequence analysis to obtain all base sequences.

[0061] The base sequence analysis of the D-loop region was performed with Perkin-Elmer DNA terminator Cycle Sequencing Ready Reaction Kit, DNA amplifier (GeneAmp PCR System 9700, Ap...

Embodiment 3

[0065] (Example 3) Preparation and confirmation of specific primers

[0066] After determining the standard D-loop base sequence of red deer, sika deer, red deer, and reindeer, FOR primer (sense; Tm value 58°C) (SEQ ID NO: 4) was used as the forward primer, and the Sika deer, red deer and reindeer specific reaction primers (serial number 5 to 8), each primer is called RED (Tmvalue 53℃), NIP (Tm value 53℃), ELK (Tm value 53℃), REIN (Tm value) 54°C). To make primers, use http: / / www.bioneer.co.kr / tools website.

[0067] In order to obtain the specific D-loop mtDNA section of the antler species, 2 pmole of FOR, RED, NIP, ELK, REIN primers and 40 pmole of L14724 (Masudaet.al., 1996) primers, 10X reaction buffer (Applied biosystems) as internal control , USA), 1.5mMMgCl2, 0.2mM dNTP Mixture, 1.25unit of AmpliTaq Gold polymerase (Applied biosystems, USA) and 1ng of DNA were added to 25μl of reaction solution.

[0068] After DNA amplification was performed at 95°C for 12 minutes in the pr...

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Abstract

The invention relates to primers specific to cervus elaphus, c. nippon, c. canadensis and rangifer tarandus gene and the method to identify cervi parvum cornu species, in particularly to method for identifying cervi parvum cornu species by use of the primers with specific sequence of cervus elaphus, c. nippon, c. canadensis and rangifer tarandus gene and a multiplex-PCR (polymerase chain reaction) of these primers. Theses primers and method using thereof to identify cervi parvum cornu species can correctly distinguish the cervi parvum cornu which is not easy to be identified in slice state and can be effectively used in prevention of illegal circulation of animal cornu other than cervi parvum cornu.

Description

Technical field [0001] The present invention relates to primers specific to red deer (Cervus elaphus), sika deer (C.nippon), red deer (C.canadensis) and reindeer (Rangifer tarandus) and methods for identifying antler species by using the primers, in particular to , Red deer and reindeer genes have primers with specific sequences and a method for identifying antler species by multiplex PCR using the primers. Background technique [0002] Antler (Cervi Parvum Cornu) is a substance obtained by cutting the unossified young horns of sika deer (Cervus nippon), a subspecies of red deer (C.elaphus L.), or red deer (C.elaphus canadensis) to dry them. Including a variety of amino acids, inorganic substances, sugars, etc. (Kim et al., 1973). [0003] Sika deer (Cervus nippon), widely distributed in the northeast (Ohtaishi 1986, Ohtaishiet. Al. 1990, Whitehead 1972, White head 1993), is slightly smaller than other deer in size and weighs 80kg, body length is 95-180cm, shoulder height is 65 ~...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12N15/11C12Q1/6844C12Q2537/143C12Q2565/125
Inventor 高柄燮吴承恩李美英金应秀金令和
Owner KOREA INST OF ORIENTAL MEDICINE
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