KR101545935B1 - Composition for identifying primate species or primate individual marker using minisatellite of SCK/SLI gene - Google Patents

Abstract

The present invention relates to a mini satellite marker composition for identification of primate or primate individuals using mini satellite of SCK / SLI gene. By confirming that the tandem repeat (TR) region in the intron region of the SCK / SLI gene exhibits the interspecific or interspecific difference according to the present invention, it is possible to identify primate identification and / or primer identification using minisatellite (MS) And can be usefully used for DNA typing.

Description

[0001] The present invention relates to a miniaturized satellite marker composition for identification of primate or primate individuals using a miniature satellite of the SCK / SLI gene,

The present invention relates to a mini satellite marker composition for identification of primate or primate individuals using mini satellite of SCK / SLI gene.

Population genetics is experiencing a dramatic development in recent years. In the 1960s, the main research area was to measure the frequency of heterozygosity and genetic polymorphism in natural populations using molecular markers such as allozyme. However, since the mid-1980s, sophisticated molecular biology techniques, including polymerase chain reaction (PCR), have been developed that have abundant mutations such as microsatellite, minisatellite and AFLP (amplified fragment length polymorphism) DNA-based molecular markers have been developed and applied. In particular, mini satellites can be genotyped by simple PCR and electrophoresis, and have very high polymorphism and are the most widely used molecular markers in population genetics. Using this, various population genetic analyzes such as individual identification, population structure and size variation are being performed.

Human SCK / SLI gene is located on chromosome (19p13.1) 19, the human genome project during repeat sequence interval is increased sequencing limit the GAP1 region of chromosome 19 present in human SCK / SLI gene was The part was also found to have many repeat sites.

In addition, repetitive sites of the SCK / SLI gene were found to have different repetition frequency and repeat units in primates such as chimpanzees, gorillas, orangutans and Lethus monkeys. Ten of these serially repetitive sites exhibited high polymorphism, But it was identified as being able to identify differences between individuals.

About 3% of the tandem repeat (TR) or varicable number of tendam repeat (VNTR) is present in the human genome, of which the length of the minisatellite (MS) is mainly 10 to 100 bp ) Repeating units and vary in length from several hundred bp to more than about 20 kb depending on the number of repeats.

The repetitive sequence is mainly present in the gene regulatory region and the intron portion, and sometimes in the exon, and may have a significant effect on the function of the gene. It is also used to understand the evolution of the whole genome and confirm the distance of the genetic evolution by showing the minute difference of the repeating unit sequence or the difference of the repeating unit depending on the species. Thus, it has been reported that the serially repetitive site plays an important role in understanding gene evolution and species evolution.

Currently, these repetitive sites are also used for the identification of paternity and forensic markers and for predicting the prognosis of diseases by exploiting the fact that they show differences between individuals or species depending on the difference in repetition unit sequence and repetition frequency. Is widely used as an essential marker to identify the marker.

Accordingly, the present inventors have completed a primate identification kit using the minisatellite (MS) of the present invention by confirming that the repeated sequence regions existing in the intron region of the SCK / SLI gene represent interspecific or individual differences.

It is an object of the present invention to provide a miniature satellite marker composition for identification of a primate or primate individual comprising at least one base sequence selected from SEQ ID NOS: 1-35.

Another object of the present invention is a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 36 and SEQ ID NO: 37; SEQ ID NO: 38 and SEQ ID NO: 39; SEQ ID NO: 40 and SEQ ID NO: 41; SEQ ID NO: 42 and SEQ ID NO: 43; SEQ ID NO: 44 and SEQ ID NO: 45; SEQ ID NO: 46 and SEQ ID NO: 47; SEQ ID NO: 48 and SEQ ID NO: 49; SEQ ID NO: 50 and SEQ ID NO: 51; SEQ ID NO: 52 and SEQ ID NO: 53; SEQ ID NO: 54 and SEQ ID NO: 55; SEQ ID NO: 56 and SEQ ID NO: 57; SEQ ID NO: 58 and SEQ ID NO: 59; SEQ ID NO: 60 and SEQ ID NO: 61; SEQ ID NO: 62 and SEQ ID NO: 63; SEQ ID NO: 64 and SEQ ID NO: 65; SEQ ID NO: 66 and SEQ ID NO: 67; SEQ ID NO: 68 and SEQ ID NO: 69; and a primer set for amplifying a miniature satellite for identifying a primate or primate.

It is still another object of the present invention to provide a probe for identifying a primate or a primate, comprising any one of the nucleotide sequences selected from SEQ ID NOS: 1 to 35, or a complementary base sequence thereof.

Yet another object of the present invention is to provide a microarray for primate or primate identification, which comprises a nucleotide consisting of any one of the nucleotide sequences selected from SEQ ID NOS: 1 to 35, a polypeptide encoded thereby, or cDNA thereof.

Another object of the present invention is to provide

1) performing PCR by mixing the primer set with all DNA samples extracted from primates;

2) DNA sequencing using the amplified DNA sample of step 1); And

3) analyzing the DNA sequencing in the step 2) and analyzing the DNA sequencing in comparison with any one of SEQ ID NOs: 1 to 35;

And a method for identifying a primate or primate object.

The present invention provides a miniature satellite marker composition for identification of a primate or primate individual comprising at least one base sequence selected from SEQ ID NOS: 1-35.

In addition, the present invention provides an isolated nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 36 and SEQ ID NO: 37; SEQ ID NO: 38 and SEQ ID NO: 39; SEQ ID NO: 40 and SEQ ID NO: 41; SEQ ID NO: 42 and SEQ ID NO: 43; SEQ ID NO: 44 and SEQ ID NO: 45; SEQ ID NO: 46 and SEQ ID NO: 47; SEQ ID NO: 48 and SEQ ID NO: 49; SEQ ID NO: 50 and SEQ ID NO: 51; SEQ ID NO: 52 and SEQ ID NO: 53; SEQ ID NO: 54 and SEQ ID NO: 55; SEQ ID NO: 56 and SEQ ID NO: 57; SEQ ID NO: 58 and SEQ ID NO: 59; SEQ ID NO: 60 and SEQ ID NO: 61; SEQ ID NO: 62 and SEQ ID NO: 63; SEQ ID NO: 64 and SEQ ID NO: 65; SEQ ID NO: 66 and SEQ ID NO: 67; SEQ ID NO: 68, and SEQ ID NO: 69; and a primer set for amplifying a miniature satellite for identifying a primate or primate object, which is composed of any one pair of primers selected from the group consisting of SEQ ID NO: 68 and SEQ ID NO: 69.

The present invention also provides a primate or primate identification probe comprising any one of the nucleotide sequences selected from SEQ ID NOS: 1-35, or a complementary base sequence thereof.

Further, the present invention provides a microarray for primate or primate identification, comprising a nucleotide consisting of any one of the nucleotide sequences selected from SEQ ID NOS: 1-35, a polypeptide encoded thereby, or a cDNA thereof.

In addition,

1) performing PCR by mixing the primer set with all DNA samples extracted from primates;

2) DNA sequencing using the amplified DNA sample of step 1); And

3) analyzing the DNA sequencing in the step 2) and analyzing the DNA sequencing in comparison with any one of SEQ ID NOs: 1 to 35;

A primate identification or a primate identification method.

By confirming that the tandem repeat (TR) region in the intron region of the SCK / SLI gene according to the present invention shows a difference between species or individuals, it is possible to use the minisatellite (MS) of the present invention Primate identification, and DNA typing.

Figure 1 shows a human SCK / SLI It is a schematic diagram of gene structure. It is a schematic diagram of the region located in the SCK / SLI gene using the sequence information of the NCBI website.
FIG. 2 is an electrophoresis photograph showing TR1 to TR7 polymorphisms in the repetitive sequence of the SCK / SLI gene. FIG.
3 is an electrophoresis photograph showing the polymorphisms of TR8 to TR12 among the repeated sequences of the SCK / SLI gene.

The present invention provides a miniature satellite marker composition for identification of a primate or primate individual comprising at least one base sequence selected from SEQ ID NOS: 1-35.

Further, the present invention relates to the aforementioned mini satellite as SEQ ID NO: 36 and SEQ ID NO: 37; SEQ ID NO: 38 and SEQ ID NO: 39; SEQ ID NO: 40 and SEQ ID NO: 41; SEQ ID NO: 42 and SEQ ID NO: 43; SEQ ID NO: 44 and SEQ ID NO: 45; SEQ ID NO: 46 and SEQ ID NO: 47; SEQ ID NO: 48 and SEQ ID NO: 49; SEQ ID NO: 50 and SEQ ID NO: 51; SEQ ID NO: 52 and SEQ ID NO: 53; SEQ ID NO: 54 and SEQ ID NO: 55; SEQ ID NO: 56 and SEQ ID NO: 57; SEQ ID NO: 58 and SEQ ID NO: 59; SEQ ID NO: 60 and SEQ ID NO: 61; SEQ ID NO: 62 and SEQ ID NO: 63; SEQ ID NO: 64 and SEQ ID NO: 65; SEQ ID NO: 66 and SEQ ID NO: 67; SEQ ID NO: 68, and SEQ ID NO: 69; and a primer set for amplifying a miniature satellite for identifying a primate or primate object, which is composed of any one pair of primers selected from the group consisting of SEQ ID NO: 68 and SEQ ID NO: 69.

Hereinafter, the present invention will be described in detail.

As used herein, the term 'primates' refers to a group of animals that form one neck of a vertebrate animal vertebrate animal mammalian mammal, and includes monkeys in the broad sense, animals that constitute a mammalian species (primate) And preferably refers to a human, chimpanzee, gorilla, orangutan or Lethus monkey, but is not limited thereto.

The term 'minisatellite' as used in the present invention refers to the repeated occurrence of 10 to 60 short nucleotide sequences. The nucleotide sequence has very high polymorphism in the number of repetitive sequences, It is recognized as a preferred marker for structural analysis. These molecular markers can be used to perform a variety of highly reliable assays by applying simple PCR techniques and utilizing a myriad of gene loci that follow Mendel's genetic rules. Therefore, it has very high discrimination power and information power compared to isozyme and protein polymorphism used in past research. It has the advantage of being able to process large amounts of data because it can semi-automate processes such as scoring and genotyping.

In the present invention, mini satellites include mini satellites in which SEQ ID NOS: 1, 4, 7, 10, 15, 19, 23, 26, 30 and 33 identify humans; SEQ ID NOS: 2, 5, 11, 16, 20, 24, 27, 31 and 34 are mini satellites identifying chimpanzees; SEQ ID NOS: 3, 6, 9, 12, 17, 21, 25, 28, 32 and 35 are mini satellites identifying gorillas; SEQ ID NOS: 13, 18, 22, and 29 are mini satellites that identify orangutans. Preferably, the kit includes a miniature satellite marker composition for primate or primate individual identification.

The term 'DNA typing' used in the present invention refers to the determination of individual identification and blood relationship using the over-variation phenomenon of the DNA base sequence and is also referred to as DNA fingerprinting. A study of genetic fingerprints began in 1985 when Jeffreys of the United Kingdom first announced that it could utilize genetically highly mutated regions of mutation, such as fingerprints, for personal identification. DNA fingerprinting, or DNA typing, is now being performed by internationally standardized analysis techniques for more useful gene markers. The gene markers used for DNA typing are mostly nonfunctional genes, and currently the most widely used markers in the world are microsatellite short tandem repeats (STRs) and mini satellites Lt; / RTI > variable number of tandem repeats (VNTRs).

As used herein, the term 'primer' refers to a nucleic acid sequence having a short free 3 'hydroxyl group, which can form a base pair with a complementary template, Refers to a short nucleic acid sequence that functions as a starting point. Primers can initiate DNA synthesis in the presence of reagents (DNA polymerase or reverse transcriptase) for polymerisation and four different dNTPs (deoxynucleoside triphosphate) at appropriate buffer solutions and temperatures.

Primers can incorporate additional features that do not alter the primer's basic properties that serve as a starting point for DNA synthesis. The primers of the present invention can be chemically synthesized using the phosphoramidite solid support method, or other well-known methods. Such nucleic acid sequences may also be modified using many means known in the art. Non-limiting examples of such modifications include, but are not limited to, methylation, "capping ", substitution of one or more natural nucleotides into homologues, and modifications between nucleotides, such as uncharged linkers such as methylphosphonate, phosphotriester, (E.g., phosphoramidate, carbamate, etc.) or charged linkages (e.g., phosphorothioate, phosphorodithioate, etc.). The nucleic acid can be in the form of one or more additional covalently linked residues such as a protein such as a nuclease, a toxin, an antibody, a signal peptide, a poly-L-lysine, an intercalator such as acridine, ), Chelating agents (e.g., metals, radioactive metals, iron, oxidizing metals, etc.), and alkylating agents.

In addition, the primer nucleic acid sequences of the present invention may, if necessary, comprise markers detectable directly or indirectly by spectroscopic, photochemical, biochemical, immunochemical or chemical means. Examples of labels include enzymes (e.g., horseradish oxidase, alkaline phosphatase), radioactive isotopes (e.g., ³² P), fluorescent molecules, chemical groups (such as biotin), and the like.

In the present invention, the primer set

SEQ ID NOS: 36 and 37; SEQ ID NOS: 42 and 43; SEQ ID NOS: 46 and 47; SEQ ID NOS: 50 and 51; SEQ ID NOS: 52 and 53; SEQ ID NOS: 54 and 55; SEQ ID NOS: 58 and 59; SEQ ID NOS: 62 and 63; And SEQ ID NOS: 64 and 65; a primer set for mini satellite amplification that identifies a human consisting of a pair of primers selected from the group consisting of SEQ ID NOS: 64 and 65;

SEQ ID NOS: 38 and 39; 40 and 41; 44 and 45; 46 and 47; 50 and 51; 52 and 53; 56 and 57; 58 and 59; 62 and 63; 66 and 67; A primer set for mini satellite amplification that identifies a chimpanzee composed of any one pair of primers selected from the group consisting of SEQ ID NOS: 68 and 69;

SEQ ID NOS: 38 and 39; 40 and 41; 44 and 45; 46 and 47; 50 and 51; 52 and 53; 56 and 57; 60 and 61; 62 and 63; 66 and 67; A primer set for mini satellite amplification that identifies a gorilla composed of any one pair of primers selected from the group consisting of SEQ ID NOS: 68 and 69; And

48 and 49; 50 and 51; 52 and 53; 58, and 59; and a primer set for mini satellite amplification that identifies orangutans composed of any one pair of primers selected from the group consisting of:

The 'primate identification PCR kit' of the present invention may include, in addition to the above primer set, conventional components included in the PCR kit. Typical components included in the kit may be reaction buffers, polymerases, dNTPs (dATP, dCTP, dGTP and dTTP), and joins such as Mg ²⁺ . A variety of DNA polymerases can be used in the amplification step of the present invention, including the Klenow fragment of E. coli DNA polymerase I, thermostable DNA polymerase, and bacteriophage T7 DNA polymerase. Preferably, the polymerase is a thermostable DNA polymerase obtainable from a variety of bacterial species, including Thermus aquaticus (Taq), Thermus thermophilus (Tth), Thermus filiformis, Thermis flavus, Thermococcus literalis, and Pyrococcus furiosus (Pfu) . Most of these enzymes can be isolated from the bacteria itself or commercially available. In addition, the polymerase used in the present invention can be obtained from cells expressing high levels of the cloning gene encoding the polymerase. When performing the polymerization reaction, it is preferable to provide the reaction vessel with an excessive amount of the components necessary for the reaction. The excess amount of the components required for the amplification reaction means an amount such that the amplification reaction is not substantially restricted to the concentration of the component.

All enzymes used in the amplification reaction may be active under the same reaction conditions. In fact, buffers make all enzymes close to optimal reaction conditions. Thus, the amplification process of the present invention can be carried out in a single reaction without changing conditions such as the addition of reactants.

The present invention also provides a primate or primate identification probe comprising any one of the nucleotide sequences selected from SEQ ID NOS: 1-35, or a complementary base sequence thereof.

The nucleotide sequences of SEQ ID NOS: 1 to 35 include SEQ ID NO: 36 and SEQ ID NO: 37; SEQ ID NO: 38 and SEQ ID NO: 39; SEQ ID NO: 40 and SEQ ID NO: 41; SEQ ID NO: 42 and SEQ ID NO: 43; SEQ ID NO: 44 and SEQ ID NO: 45; SEQ ID NO: 46 and SEQ ID NO: 47; SEQ ID NO: 48 and SEQ ID NO: 49; SEQ ID NO: 50 and SEQ ID NO: 51; SEQ ID NO: 52 and SEQ ID NO: 53; SEQ ID NO: 54 and SEQ ID NO: 55; SEQ ID NO: 56 and SEQ ID NO: 57; SEQ ID NO: 58 and SEQ ID NO: 59; SEQ ID NO: 60 and SEQ ID NO: 61; SEQ ID NO: 62 and SEQ ID NO: 63; SEQ ID NO: 64 and SEQ ID NO: 65; SEQ ID NO: 66 and SEQ ID NO: 67; A nucleotide sequence amplified by any one primer pair selected from the group consisting of SEQ ID NO: 68 and SEQ ID NO: 69 is preferable, but not limited thereto.

In the present invention, 'probe' refers to a hybridization probe, and refers to an oligonucleotide capable of binding sequence-specifically to the complementary strand of a nucleic acid. By using this, the genomic DNA of the primate can be extracted, and any one of the nucleotide sequences selected from SEQ ID NOS: 1 to 35 or a complementary base sequence thereof can be hybridized with the genome DNA of the primate with a probe.

The nucleotide sequence of SEQ ID NOS: 1 to 35 is a polymorphic sequence. A polymorphic sequence refers to a sequence comprising a polymorphic site representing a SNP in a polynucleotide sequence. The polynucleotide may be DNA or RNA.

The present invention also provides a primate-like primate individual microarray comprising a nucleotide consisting of any one of the nucleotide sequences selected from SEQ ID NOS: 1-35, a peptide encoded thereby, or a cDNA thereof.

The microarray according to the present invention can be produced by a general method known to a person skilled in the art.

For example, the nucleotide may be immobilized on a substrate coated with an activator selected from the group consisting of aminosilane, poly-L-lysine and aldehyde. In addition, the substrate may be selected from the group consisting of a silicon wafer, glass, quartz, metal, and plastic. The method of immobilizing the polynucleotide on a substrate may be a micropipetting method using a piezoelectric method or a method using a pin-type spotter.

In addition,

1) SEQ ID NO: 36 and SEQ ID NO: 37; SEQ ID NO: 38 and SEQ ID NO: 39; SEQ ID NO: 40 and SEQ ID NO: 41; SEQ ID NO: 42 and SEQ ID NO: 43; SEQ ID NO: 44 and SEQ ID NO: 45; SEQ ID NO: 46 and SEQ ID NO: 47; SEQ ID NO: 48 and SEQ ID NO: 49; SEQ ID NO: 50 and SEQ ID NO: 51; SEQ ID NO: 52 and SEQ ID NO: 53; SEQ ID NO: 54 and SEQ ID NO: 55; SEQ ID NO: 56 and SEQ ID NO: 57; SEQ ID NO: 58 and SEQ ID NO: 59; SEQ ID NO: 60 and SEQ ID NO: 61; SEQ ID NO: 62 and SEQ ID NO: 63; SEQ ID NO: 64 and SEQ ID NO: 65; SEQ ID NO: 66 and SEQ ID NO: 67; A primer set consisting of any one primer pair selected from the group consisting of SEQ ID NO: 68 and SEQ ID NO: 69, and a whole DNA sample extracted from the primate;

2) DNA sequencing using the amplified DNA sample of step 1); And

3) analyzing the DNA sequencing in the step 2) and analyzing the DNA sequencing in comparison with any one of SEQ ID NOs: 1 to 35;

A primate or primate entity.

The method of identifying primate or primate individuals of the present invention will be described in detail as follows.

In the step 1), the entire DNA extracted from the primate is subjected to a PCR reaction, and the term " PCR reaction " means a reaction for amplifying a nucleic acid molecule. A variety of amplification reactions have been reported in the art, including polymerase chain reaction (PCR) (US Pat. Nos. 4,683,195, 4,683,202, and 4,800,159), reverse transcription-polymerase chain reaction (RT- , The method of Miller, HI (WO 89/06700) and Davey, C. et al (EP 329,822), the method of ligase chain reaction (Sambrook et al., Molecular Cloning, A Laboratory Manual, 3rd ed. Cold Spring Harbor Press LCR (17, 18), Gap-LCR (WO 90/01069), repair chain reaction (EP 439,182), transcription-mediated amplification (TMA) WO 88/10315), self sustained sequence replication (20) (WO 90/06995), selective amplification of target polynucleotide sequences (US Pat. No. 6,410,276) , Consensus sequence primer polymerase chain reaction (CP-PCR) ( U.S. Patent No. 4,437,975), arbitrarily primed polymerase chain reaction (AP-PCR) (U.S. Patent Nos. 5,413,909 and 5,861,245), nucleic acid sequence based amplification (NASBA) (U.S. Patent Nos. 5,130,238, 5,409,818, 5,554,517, and 6,063,603), strand displacement amplification (21,22) and loop-mediated isothermal amplification (LAMP) 23, but is not limited thereto. Other amplification methods that may be used are described in U.S. Patent Nos. 5,242,794, 5,494,810, 4,988,617 and U.S. Patent No. 09 / 854,317.

The step 2) is a step of sequencing DNA using the amplified DNA. As a method for determining DNA sequence (order of arrangement of adenine, cytosine, guanine, thymine) which is widely used in DNA sequencing, there is a chemical analysis method -Gilbert method) and a chain termination method (chain termination method, Sanger method), but the method is not limited to this, and is well known in the art.

The step 3) is a step of comparing DNA sequencing analysis results. The mini satellite DNA marker amplified using the primer pair of the present invention is analyzed by a suitable method. For example, the result of the amplification reaction is subjected to gel electrophoresis, and the resulting band is observed and analyzed to analyze the characteristics of the amplified mini satellite DNA marker.

In the present invention, a fragment of the amplified mini satellite DNA marker is analyzed to determine its genotype. For example, by analyzing the size of the amplification product that varies depending on the number of repeats of the miniature satellite DNA marker, various genotypes are classified and primates are identified based on the statistical data of the classified genotypes. In other words, accumulating information on the genotypes of mini satellite DNA markers that appear or appear at high frequency in specific primates, and comparing the genotypes obtained from the samples to be identified based on the accumulated information with existing genotyping information, It is possible to identify individuals or populations.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

[Example 1] Structural analysis of SCK / SLI gene, analysis of polymorphism of minisatellite (MS) in SCK / SLI gene and analysis of tandem repeat (TR)

Structural features of SCK / SLI gene were analyzed using NCBI BLAST SEARCH. After examining the exon and intron regions and determining the location of the tandem repeat (TR) in each region, the position of the minisatellite (MS) with a repeating sequence size of about 10-100 bp Respectively. As a result, the positions including 12 TR regions in the intron region in the SCK / SLI gene were confirmed.

Of the 12 TRs identified above, SCK / SLI Polymorphisms of 10 tandem repeats (TR) in the gene were analyzed by polymerase chain reaction (PCR). The results are shown in Table 1. Also, a schematic diagram of the human SCK / SLI gene structure is shown in Fig.

Minisatellite Repeating sequence Repeat size (bp) Copies
(copy no.) TR1 H CCTCCAGCTC CTGGTCCTGG GGGGTCCTCC
CGCCCTGACC CTCACTCCCT GGTCTCCCGC C (SEQ ID NO: 1) 61 5.8 C CCTCCAGCTC CTGGTCCTGG GGGGTCCTCC
CGCCCTGACC CTCACTCCCT GGTCTCCCGC C (SEQ ID NO: 2) 61 3.7 G CCTCCAGCTC CTGGTCCTGG GGGGTCCTCC
CGCCCTGACC CTCACTCCCT GGTCTCCCGC C (SEQ ID NO: 3) 61 2.7 TR2 H AGAGTCCGGG GAGGGAGGAC GACACCGAGA GGGGC (SEQ ID NO: 4) 35 14.6 C AGAGTCCGGG GAGGGAGGAC GACACTGAGA GACGC (SEQ ID NO: 5) 35 1.0 G AGAGTCCGGG GAGGGAGGAC GACACCGAGA GGGGC (SEQ ID NO: 6) 35 6.7 TR3 H GGGGAACTGC ACACGGCACA GGGAA (SEQ ID NO: 7) 25 16.2 C GGGGAATTGC ACACGGCACA GGGAA (SEQ ID NO: 8) 25 17.5 G GGGGAATTGC GCACGGCACA GGGAA (SEQ ID NO: 9) 25 30.9 TR4 H ACCTAATACC GTGTGGATGA CGCAGTACCT ATACCCAACA TGCA-GGAA (SEQ ID NO: 10) 48 36.1 C ACCTAATACC GTGTGGATGA CGCAGTACCT ATACCCAACA TGCACAGAA (SEQ ID NO: 11) 49 29.9 G ACCTAATACC GTGTGGATGA CGCAGTACCT
ATACCCAACA TGCACGGAA (SEQ ID NO: 12) 49 36.1 O ACCTAATACC GTGTGGATGA CGCAGTACCT
ATATCCAACA TGCACGGAA (SEQ ID NO: 13) 49 33.7 R ACCTGATACC GTGTGGATGA CGCAGCACCT
ACATCCAACA CGCACGGAA (SEQ ID NO: 14) 49 22.4 TR7 H ATCTGCGTGC TCGTTTGCGT GCTC (SEQ ID NO: 15) 24 7.7 C ATCTGCGTGC TCGTCTGCGT GCTC (SEQ ID NO: 16) 24 32.9 G ATCTGCATGC TCATTTGCAT GCTC (SEQ ID NO: 17) 24 7.45 O ATCTGCATGC TTGTTTGCGT GCTC (SEQ ID NO: 18) 24 14.6 TR8 H GTCATATTTC AGCGTGTGGA TGGCCACGCC GTGGCCAC
(SEQ ID NO: 19) 38 16.6 C GTCATATTTC AGCGTGTGGA TGGCCACGCC GTGGCCAC
(SEQ ID NO: 20) 38 12.6 G GTCGTATTTC AGCGTGTGGA TGGCCACGCC ATGGCCAC
(SEQ ID NO: 21) 38 17.2 O ACCATATTTC AGCGTGTGGA TGGCCACGCC ATGGCCAC
(SEQ ID NO: 22) 38 4.7 TR9 H TGCGTTTCTC CGTTTCATTG AGGTTGGGGC
ATCGTACTGA CTTGGGGGGA ATTCCTGCGT AGGTG
(SEQ ID NO: 23) 65 7.3 C TGCGTTTCTC CGTTTCATTG AGGTTGGGGC
ATCGTACTGA CTTGGGGG-A GTTCCTGCGT AGGTG
(SEQ ID NO: 24) 64 23.5 G TGTGTTTCTC CGTTTCATTG AGGTTGGGGC
ATCGTACTGA CTTGGGGG-A GTTCCTGCGT AGGTG
(SEQ ID NO: 25) 64 5.7 TR10 H TGCACCTGCT GTGCCCCGCC TAGAACTCTG
TCTGCAAACC CCACCACGTC AGGCCTTTGC ACC (SEQ ID NO: 26) 63 12.4 C TGCACCTGCT GTGCCCCGCC TAGAACTCTG
TCTGCAAACC C-ACCACGTC AGGCCTTTGC ACC (SEQ ID NO: 27) 62 8.5 G TGCACCTGCT GTCCCCC-CC TAGAACTCTG
TCTGCAAACT CCACCACGTC AGGCCTTTGC ACC (SEQ ID NO: 28) 62 8.2 O TGCACCTGCT GTGCCCCGCC TAGAACTCTG
TCTGCAAACT CCACCGCATC GGGCCTTTCC ACC (SEQ ID NO: 29) 63 6.0 TR11 H GGGGAGGCGG AAGCGGGTCC C (SEQ ID NO: 30) 21 44.6 C GGGGAGGCGG AAGCGGGTCC C (SEQ ID NO: 31) 21 34.0 G GGGGAGGCGG AAGCGGGTCC T (SEQ ID NO: 32) 21 23.7 TR12 H GTAGGGGGAA GGACCCCACT GAACCCAGC (SEQ ID NO: 33) 29 16.8 C GTAGGGGGAA GGACCCCACT GAACTCAGT (SEQ ID NO: 34) 29 15.3 G GTAGGGGGAA GGACCCCACT GAACCCAGC (SEQ ID NO: 35) 29 9.1

SCK / SLI The nucleotide sequences of the primers used in the PCR analysis of the 10 serial repeats in the gene are shown in Table 2 below.

Minisatellite  Base sequence Bell TR1 F GGCGGCCTTGAATTCCTCCA
(SEQ ID NO: 36) R CTCAGCCCCCAGGGTCAGG
(SEQ ID NO: 37) H F TGACAGGCCACTCTAAGGCTCTCC
(SEQ ID NO: 38) R ATGCTGACTGGCCAGGCTAGGAGTT
(SEQ ID NO: 39) C, G TR2 F GGTCAGGATGGGTGAGAGGGAAG
(SEQ ID NO: 40) R CCTGCGTCTCGCACTGTCGT
(SEQ ID NO: 41) H, C, G TR3 F ACCTGAGGGAGCGGGAAGGA
(SEQ ID NO: 42) R TGCCATGTGGCCTCCCTTTT
(SEQ ID NO: 43) H F GATCTCAATGCCCATCAGGAATG
(SEQ ID NO: 44) R CTGGGCTCAAGAGATCCTCCCTTC
(SEQ ID NO: 45) C, G TR4 F CCATGATGTTTTGAACCATGTGGA
(SEQ ID NO: 46) R AGATCGTCTTGCTCCTAACACTCTGC
(SEQ ID NO: 47) H, C, G, R F GGGAGGCAAAGAAAACAGATGCAG
(SEQ ID NO: 48) R GCTGCAAGGGGTACCCACGA
(SEQ ID NO: 49) O TR7 F GAAGTCATGTCTGAAATGCCACCTC
(SEQ ID NO: 50) R CACAATGGGGCGCCGTGTAT
(SEQ ID NO: 51) H, C, G, O TR8 F TTCTTTCCTTGGCCGCATCA
(SEQ ID NO: 52) R CCAGCAGCCCAAAGGTGGAA
(SEQ ID NO: 53) H, C, G, O TR9 F CAGGAATGGTGGCCTTTGTGG
(SEQ ID NO: 54) R CCCACCTACGCAGGAACTCCA
(SEQ ID NO: 55) H F GGGAGGCAAAGAAAACAGATGCAG
(SEQ ID NO: 56) R GCTGCAAGGGGTACCCACGA
(SEQ ID NO: 57) C, G  TR10 F CCCGCTCCCTCTCCATCAG
(SEQ ID NO: 58) R GCCGTGGGGTTTGCAGACAT
(SEQ ID NO: 59) H, C, O F ACACTCTGCGCCACCCCCTA
(SEQ ID NO: 60) R CTCCCAGGAGGCGACCTTGA
(SEQ ID NO: 61) G  TR11 F AGGACAAACCGCCGGGAGAC
(SEQ ID NO: 62) R AGCCCCCAGAAGCTGGCT
(SEQ ID NO: 63) H, C, G  TR12 F TGCAAGGACCCTGTTCCCAAT
(SEQ ID NO: 64) R GGTGGGTGCTCCCAGCTCCT
(SEQ ID NO: 65) H F AGAGACCCCTGATCCAGGTAGGC
(SEQ ID NO: 66) R TGCGGCCAGGAACATTAGCC
(SEQ ID NO: 67) C, G F ACAGCCAGCTTCTGGGGGCT
(SEQ ID NO: 68) R TGCGGCCAGGAACATTAGCC
(SEQ ID NO: 69) C, G

Using genomic DNA extracted from blood of 103 adult males and females, TR1 located in intron 10 region, TR2 located in intron 9 region, and intron 9 region in 12 TRs present in SCK / SLI TR4 located in the intron 9 region, TR5 located in the intron 8 region, TR6 located in the intron 8 region, TR7 located in the intron 4 region, TR8 located in the intron 1 region, TR1 located in the intron 1 region, TR11 located in the intron 1 region, and TR12 located in the intron 1 region. TR1, TR2, TR3, TR4, TR7, TR8 , TR9, TR10, TR11, and TR12 were compared using PCR.

1. PCR experiment conditions of human genome DNA

Standard PCR Conditions Genomic DNA (50 mM Tris-HCl ( pH 9.0), 50 mM MgCl 2, 0.2 mM dTTP, 0.2 mM dCTP, 0.2 mM dGTP, 0.2 mM dATP, final volume 50 ㎕ - desired make any base unit) TR1 is a primer pair of SEQ ID NOs: 36 and 37, TR2 is a primer pair of SEQ ID NOs: 40 and 41, TR3 is a primer pair of SEQ ID NOs: 42 and 43, TR4 is a primer pair of SEQ ID NOs: 46 and 47, TR7 is a primer pair of SEQ ID NO: And TR8 is a pair of primers of SEQ ID NOS: 52 and 53, TR9 is a pair of primers of SEQ ID NOS: 54 and 55, TR10 is a pair of primers of SEQ ID NOS: 58 and 59, TR11 is a pair of primers of SEQ ID NOS: 62 and 63, TR12 was amplified using the primer pairs of SEQ ID NOS: 64 and 65.

PCR analysis of the DNA samples was performed using GoTaq Flexi DNA polymerase with 100 ng of genomic DNA as a template. The conditions of the PCR cycle were as follows: TR1 and TR3 were heat treated once at 94 DEG C for 2 minutes, followed by 30 cycles of 45 seconds at 94 DEG C, 30 seconds at 60 DEG C and 45 seconds at 72 DEG C, Lt; RTI ID = 0.0 > 72 C < / RTI > for 7 minutes. For TR2, 30 cycles of 45 seconds at 94 ° C, 30 seconds at 55 ° C, and 45 seconds at 72 ° C were performed after one heat treatment at 94 ° C for 2 minutes, and then the last elongation step was elongated at 72 ° C for 7 minutes . In the case of TR4, TR8, TR9, TR10, TR11 and TR12, after one heat treatment at 94 DEG C for 2 minutes, followed by 30 cycles of 94 DEG C for 45 seconds and 68 DEG C for 2 minutes, Lt; / RTI > for 7 min. For TR7, 30 cycles of 94 ° C for 30 seconds, 57 ° C for 30 seconds, and 72 ° C for 30 seconds were performed after one heat treatment at 94 ° C for 2 minutes, and then the last extension step was elongated at 72 ° C for 7 minutes . The PCR product was injected into 1-2% SeaKem LE agarose gel and analyzed by electrophoresis (1 volt / cm) in TAE buffer. As a result, all of the TR regions showed polymorphic satellites, and thus it could be used as a personal identification marker. Therefore, additional analysis was conducted by increasing the number of individuals. N is the number of alleles, which is twice the number of samples since it has two alleles for each individual.

2. Allelopathological analysis of TR part of human SCK / SLI gene

(1) TR1 (SEQ ID NO: 1)

A total of 103 alleles of 130 bp, 190 bp, 250 bp, 310 bp, 370 bp, 420 bp, 540 bp, 615 bp and 720 bp were identified from 103 normal individuals, indicating that the 61 bp repeating unit was 1.7 , 2.7, 3.7, 4.7, 5.7, 6.7, 8.7, 9.7 and 11, respectively (Table 1, Fig. 2).

(2) TR2 (SEQ ID NO: 4)

Seventy alleles of 400 bp, 500 bp, 540 bp, 570 bp, 600 bp, 640 bp and 680 bp were found in the normal subjects, indicating that the 35 bp repeating units were 9.7, 12.7, 13.7 and 14.7 , 15.7, 16.7 and 17.7 times, respectively (Table 1, Fig. 2).

(3) TR3 (SEQ ID NO: 7)

Five normal alleles of 370 bp, 400 bp, 430 bp, 460 bp, and 560 bp were identified in 103 normal individuals, indicating that 25 bp repeat units were repeated at 12.9, 13.9, 14.9, 15.9 and 18.9 times Respectively (Table 1, Fig. 2).

(4) TR4 (SEQ ID NO: 10)

Four normal alleles of 1.8 kb, 1.85 kb, 2.3 kb, and 2.45 kb in size were identified in the study of 103 normal individuals, with 48 bp repeat units of 34.3, 35.3, 44.3, and 47.3 repeats, respectively 1, Fig. 2).

(5) TR7 (SEQ ID NO: 15)

In a study of 103 normal individuals, it was confirmed that the formation of 410 bp was a repetitive unit of 24 bp, 7.7 repeats (Table 1, Fig. 2).

(6) TR8 (SEQ ID NO: 19)

Two normal alleles of 680 bp and 720 bp were identified as 103 bp normal individuals, and the 38 bp repeats were repeated 15 times and 16 times, respectively (Table 1, Fig. 3).

(7) TR9 (SEQ ID NO: 23)

Three normal alleles of 410 bp, 550 bp and 620 bp were identified in 103 normal individuals, with 38 bp repeats being 4.7, 6.7, and 7.7 repeats, respectively (Table 1, Figure 3) .

(8) TR10 (SEQ ID NO: 26)

Four normal alleles were identified as 740 bp, 800 bp, 860 bp, and 1070 bp in size, and the 62 bp repeats were 11, 11.7, 12.7, and 16 repeated, respectively 1, Fig. 3).

(9) TR11 (SEQ ID NO: 30)

Four normal alleles were identified as 1030 bp, 1060 bp, 1080 bp, and 1100 bp in size, and the repeat units of 21 bp were 41.7, 42.7, 43.7, and 44.7 repeated times, respectively 1, Fig. 3).

(10) TR12 (SEQ ID NO: 33)

Three normal alleles of 670 bp, 730 bp, and 760 bp were identified in 103 normal individuals, with 29 bp repeat units of 16.8, 18.8, and 19.8 repeat sizes, respectively (Table 1, Figure 3) .

[Example 2] Identification of a serial repeating section using primate genomic DNA

The genomic DNA of human primates belongs to genomic DNA of chimpanzees, gorillas, orangutans and Lethus monkeys. Thus, the genomic DNA of human, chimpanzee, gorilla, orangutan and Lethus monkeys consists of a number of similar nucleotide sequences, but it can be used as a measure of molecular biological evolution, It is possible to do.

Thus, in the present invention, PCR was carried out in the same manner as in Example 1 using human genomic DNA and genomic DNAs of chimpanzee, gorilla, orangutan, lysus and monkey, whereby allelic patterns of the TR portion of the SCK / SLI gene Respectively.

Using 5 genomic DNAs extracted from chimpanzee cells, 2 genomic DNAs extracted from gorilla cells, 5 genomic DNAs extracted from orangutan cells, and 13 genomic DNAs extracted from Rhesus monkey cells, 12 TRs present in SCK / SLI Allelopathic patterns of 10 parts of TR1, TR2, TR3, TR4, TR7, TR8, TR9, TR10, TR11 and TR12 were compared using PCR. Under the standard PCR conditions (50 mM Tris-HCl (pH 9.0), 50 mM MgCl 2, 0.2 mM dTTP, 0.2 mM dCTP, 0.2 mM dGTP, 0.2 mM dATP, final volume 50 μl) 39, TR2 is a primer pair of SEQ ID NOs: 40 and 41, TR3 is a primer pair of SEQ ID NOs: 44 and 45, TR4 is a primer pair of SEQ ID NOs: 48 and 49, TR7 is a primer pair of SEQ ID NOs: 50 and 51, TR8 TR12 is a primer pair of SEQ ID NOS: 62 and 63, TR12 is a primer pair of SEQ ID NOS: 62 and 63, TR12 is a primer pair of SEQ ID NOS: Or a pair of primers of SEQ ID NOS: 68 and 69, respectively. PCR analysis of the DNA samples was performed using GoTaq Flexi DNA polymerase with 100 ng of genomic DNA as a template.

The conditions of the PCR cycle are specified below for the differences between the apes.

1. PCR experiment conditions of primate genomic DNA

(1) PCR experiment conditions of chimpanzee

In the case of TR1, TR3, TR4, TR7, TR8, TR9, TR10, TR11 and TR12, the PCR experiment conditions of the human genomic DNA were carried out in the same manner as in Example 1 above.

For TR2, 30 cycles of 45 seconds at 94 ° C, 30 seconds at 56 ° C, and 45 seconds at 72 ° C were performed after one heat treatment at 94 ° C for 2 minutes, and then the last elongation step was elongated at 72 ° C for 7 minutes Respectively.

(2) PCR conditions of gorilla

In the case of TR1, TR2, TR3, TR7, TR8, TR9, TR10 and TR12, the PCR experiment conditions of the human genomic DNA were carried out in the same manner as in Example 1 above.

For TR4, after one heat treatment at 94 ° C for 2 minutes, followed by 30 cycles of 94 ° C for 45 seconds and 69 ° C for 2 minutes, the last extension step was elongated at 72 ° C for 7 minutes.

In the case of TR11, 30 cycles of 94 ° C for 30 seconds, 61 ° C for 10 seconds, and 72 ° C for 1 minute were performed after one heat treatment at 94 ° C for 2 minutes, followed by a final extension step at 72 ° C for 7 minutes Lt; / RTI >

(3) PCR experiment conditions of orangutan

TR4 and TR7 were carried out in the same manner as the PCR experiment conditions of human genomic DNA in Example 1 above.

In the case of TR8, 30 cycles of 94 ° C for 30 seconds, 57 ° C for 30 seconds, and 72 ° C for 20 seconds were performed after one heat treatment at 94 ° C for 2 minutes, and then the last extension step was performed at 72 ° C for 7 minutes Lt; / RTI >

For TR10, 30 cycles of 94 ° C for 30 seconds, 55 ° C for 15 seconds, and 72 ° C for 45 seconds were performed after one heat treatment at 94 ° C for 2 minutes, and then the last elongation step was performed at 72 ° C for 7 minutes Respectively.

(4) PCR experiment conditions of Lethus monkey

For TR4, 30 cycles of 94 ° C for 30 seconds, 55 ° C for 15 seconds, and 72 ° C for 45 seconds were performed after one heat treatment at 94 ° C for 2 minutes, and then the last elongation step was performed at 72 ° C for 7 minutes Respectively.

The PCR product was injected into 1-2% SeaKem LE agarose gel and analyzed by electrophoresis (1 volt / cm) in TAE buffer. As a result, some TR regions showed polymorphic satellites, and thus it could be used as an object identification marker.

Therefore, an allelic aspect analysis of the TR part was conducted below. N is the number of alleles, which is twice the number of samples since it has two alleles for each individual.

2. Allelopathological analysis of the TR part of the SCK / SLI gene in primates

(1) Allelopathological analysis of chimpanzee TR

1) TR1

A total of 250 bp alleles were identified from 5 chimpanzee individuals, with a repeat size of 61 bp repeated 3.7 times (Table 1, Fig. 2).

2) TR2

A total of 160 bp alleles were identified from 5 chimpanzees, with 35 bp repeats being 1.7 repeats (Table 1, Figure 2).

3) TR3

Four alleles of 280 bp, 430 bp, 460 bp, and 710 bp were identified as 5 repeated chimpanzees, and the 25 bp repeats were 9.9, 14.9, 15.9 and 23.9 times (Table 1, Fig. 2).

4) TR4

Three alleles of 1.15 kb, 1.6 kb, and 1.65 kb in size were identified in 5 chimpanzees, with 49 bp repeat units of 28.3, 30.3, and 31.3 repeats, respectively (Table 1, 2).

5) TR7

Three alleles of 986 bp, 1034 bp, and 1106 bp in size were found in the chimpanzee, and the repeat units of 24 bp were 31.7 times, 33.7 times, and 36.7 times, respectively (Table 1 2).

6) TR8

Three alleles of 490 bp and 530 bp in size were identified in 5 chimpanzees, and the 38 bp repeats were 11 and 12 repeated, respectively (Table 1, Fig. 3).

7) TR9

Five alleles of 1320 bp, 1670 bp, 1740 bp, 1810 bp and 1880 bp were identified as a result of the examination of 5 chimpanzees, and the 64 bp repeating units were identified as 17.7, 22.7, 23.7, 24.7, 25.7 (Table 1, Fig. 3).

8) TR10

As a result of examination of 5 chimpanzees, two alleles of 620 bp and 660 bp size were identified, and 62 bp repeating units were 7.7 times and 8.7 times repeated, respectively (Table 1, Fig. 3).

9) TR11

Three alleles of 840 bp, 860 bp and 880 bp were identified in the chimpanzee, and the repeat units of 21 bp were 32.7, 33.7 and 34.7 times, respectively (Table 1, Fig. 3 ).

10) TR12

Three alleles of 560 bp, 610 bp, and 700 bp in size were identified in 5 chimpanzees, and the 29 bp repeats were 12.8, 14.8, and 17.8 times repeated (Table 1, Figure 3) .

(2) Allelopathological analysis of TR part of gorilla

1) TR1

A single 190 bp allele of the gorilla was identified, with a repeat size of 61 bp repeated 2.7 times (Table 1, Figure 2).

2) TR2

Two alleles of 250 bp and 350 bp in size were observed in the gorilla (Fig. 1, Fig. 2). These results indicate that the 35 bp repeats were repeated 5.7 times and 7.7 times, respectively.

3) TR3

Four alleles were identified as 770 bp, 920 bp, 980 bp, and 1010 bp, which were repeated 25.9 times, 30.9 times, 32.9 times and 33.9 times, respectively. (Table 1, Fig. 2).

4) TR4

Two alleles of 1.15 kb and 1.30 kb in size were identified, and 49 bp repeats were repeated 21.3 times and 25.3 times, respectively (Table 1, Fig. 2).

5) TR7

Two alleles of 386 bp and 410 bp were identified in the gorilla (Fig. 1, Fig. 3). These results indicate that 24 bp repeats were repeated 6.7 times and 7.7 times, respectively.

6) TR8

Three alleles of 680 bp, 720 bp, and 870 bp were identified in the gorilla, and the 38 bp repeats were 15, 16, and 19 repeated, respectively (Table 1, 3).

7) TR9

One allele of the 480 bp size was identified as a result of the examination of 2 gorillas, and the 38 bp repeats were repeated 5.7 times (Table 1, Fig. 3).

8) TR10

Two alleles of 620 bp and 660 bp in size were observed in the gorilla (Fig. 1, Fig. 3). These results indicate that 62 bp repeats were repeated 7.7 times and 8.7 times, respectively.

9) TR11

Two alleles of 451 bp and 840 bp were observed in the gorilla (Fig. 1, Fig. 3). These results indicate that 21 bp repeats were repeated 14.7 times and 32.7 times, respectively.

10) TR12

Two alleles of 370 bp and 470 bp were identified in the gorilla (Fig. 1, Fig. 3). These results indicate that 29 bp repeats were repeated 6.8 and 9.8 times, respectively.

(3) Allelopathological analysis of the TR part of the orangutan

1) TR4

Five alleles of 1.65 kb, 1.70 kb, 1.75 kb, 1.8 kb, and 1.85 kb were identified as the result of investigation of 5 orangutans, and it was confirmed that the 49 bp repeat units were 31.3, 32.3, 33.3, 34.3, 35.3 (Table 1, Fig. 3).

2) TR7

As a result of examining five orangutans, two alleles having a size of 554 bp and 578 bp were confirmed, which are the repeated sizes of 24 bp and 13.7 times and 14.7 times, respectively (Table 1 and Fig. 3).

3) TR8

As a result of examining five orangutans, one allele of 350 bp was identified, which was repeated four times in 38 bp repeats (Table 1, Fig. 3).

4) TR10

As a result of examining five orangutans, four alleles of 580 bp, 620 bp, 660 bp, and 800 bp were identified. The 63 bp repeats were repeated 2.7 times, 7.7 times, 8.7 times, and 11.7 times (Table 1, Fig. 3).

(4) Allelopathological analysis of the TR part of Lethus monkey

1) TR4

Three alleles of 0.5 kb, 1.3 kb and 1.35 kb in size were identified in 13 resuscus monkeys, with 49 bp repeats of 8.3, 25.3 and 26.3 repeats, respectively (Table 1 , Fig. 2).

3. Analysis of genetic polymorphism within the ape species of the TR part of the SCK / SLI gene in primates

Based on the above results, a polymorphism analysis was carried out to confirm the genetic polymorphism of the SCK / SLI gene in the ape species. Through the above analysis, it was confirmed whether or not the marker was available for the object identification. The results are shown in Table 3 below.

Minisatellite Heterozygosity Human Chimpanzee Gorilla Orangutan Rhesus macaque TR1 0.746 0.000 0.000 - - TR2 0.717 0.000 0.375 - - TR3 0.671 0.700 0.750 - - TR4 0.186 0.560 0.375 0.780 0.636 TR7 0.000 0.460 0.375 0.219 - TR8 0.100 0.180 0.625 0.000 - TR9 0.279 0.780 0.000 - - TR10 0.351 0.560 0.500 0.656 - TR11 0.076 0.540 0.500 - - TR12 0.039 0.620 0.375 - -

As shown in Table 3, according to the polymorphism analysis, in case of TR1, human. Polymorphism results of 0.746, 0.000 and 0.000 in chimpanzees and gorillas were confirmed, respectively.

In the case of TR2, polymorphism results of 0.717, 0.000 and 0.375 were found in human, chimpanzee and gorilla, respectively.

In the case of TR3, polymorphisms of 0.671, 0.700 and 0.750 in human, chimpanzee and gorilla were confirmed, respectively.

In the case of TR4, polymorphism results of 0.186, 0.560, 0.375, 0.780 and 0.636 were found in human, chimpanzee, gorilla, orangutan and Lethus monkey, respectively.

In the case of TR7, polymorphisms of 0.000, 0.460, 0.375 and 0.219 were found in humans, chimpanzees, gorillas and orangutans, respectively.

In the case of TR8, polymorphisms of 0.100, 0.180, 0.625 and 0.000 were found in humans, chimpanzees, gorillas and orangutans, respectively.

In the case of TR9, polymorphisms of 0.279, 0.780 and 0.000 were found in human, chimpanzee and gorilla, respectively.

In the case of TR10, polymorphism results of 0.351, 0.560, 0.500 and 0.656 were found in human, chimpanzee, gorilla and orangutan, respectively.

In the case of TR11, polymorphisms of 0.076, 0.540 and 0.500 were found in human, chimpanzee and gorilla, respectively.

In the case of TR12, polymorphisms of 0.039, 0.620 and 0.375 were found in human, chimpanzee and gorilla, respectively.

Through the results of the polymorphism analysis in the species of apes, it was confirmed that the miniaturized satellite marker for primate identification of the present invention can be used to identify individuals in apes.

<110> Dong-A University Research Foundation For Industry-Academy Cooperation <120> Composition for identifying primate species or primate individual          marker using minisatellite of SCK / SLI gene <130> Donga 1.45P <160> 69 <170> Kopatentin 2.0 <210> 1 <211> 61 <212> DNA <213> human SCK / SLI gene minisatellite <400> 1 cctccagctc ctggtcctgg ggggtcctcc cgccctgacc ctcactccct ggtctcccgc 60 c 61 <210> 2 <211> 61 <212> DNA <213> human SCK / SLI gene minisatellite <400> 2 cctccagctc ctggtcctgg ggggtcctcc cgccctgacc ctcactccct ggtctcccgc 60 c 61 <210> 3 <211> 61 <212> DNA <213> human SCK / SLI gene minisatellite <400> 3 cctccagctc ctggtcctgg ggggtcctcc cgccctgacc ctcactccct ggtctcccgc 60 c 61 <210> 4 <211> 35 <212> DNA <213> human SCK / SLI gene minisatellite <400> 4 agagtccggg gagggaggac gacaccgaga ggggc 35 <210> 5 <211> 35 <212> DNA <213> human SCK / SLI gene minisatellite <400> 5 agagtccggg gagggaggac gacactgaga gacgc 35 <210> 6 <211> 35 <212> DNA <213> human SCK / SLI gene minisatellite <400> 6 agagtccggg gagggaggac gacaccgaga ggggc 35 <210> 7 <211> 25 <212> DNA <213> human SCK / SLI gene minisatellite <400> 7 ggggaactgc acacggcaca gggaa 25 <210> 8 <211> 25 <212> DNA <213> human SCK / SLI gene minisatellite <400> 8 ggggaattgc acacggcaca gggaa 25 <210> 9 <211> 25 <212> DNA <213> human SCK / SLI gene minisatellite <400> 9 ggggaattgc gcacggcaca gggaa 25 <210> 10 <211> 48 <212> DNA <213> human SCK / SLI gene minisatellite <400> 10 acctaatacc gtgtggatga cgcagtacct atacccaaca tgcaggaa 48 <210> 11 <211> 49 <212> DNA <213> human SCK / SLI gene minisatellite <400> 11 acctaatacc gtgtggatga cgcagtacct atacccaaca tgcacagaa 49 <210> 12 <211> 49 <212> DNA <213> human SCK / SLI gene minisatellite <400> 12 acctaatacc gtgtggatga cgcagtacct atacccaaca tgcacggaa 49 <210> 13 <211> 49 <212> DNA <213> human SCK / SLI gene minisatellite <400> 13 acctaatacc gtgtggatga cgcagtacct atatccaaca tgcacggaa 49 <210> 14 <211> 49 <212> DNA <213> human SCK / SLI gene minisatellite <400> 14 acctgatacc gtgtggatga cgcagcacct acatccaaca cgcacggaa 49 <210> 15 <211> 24 <212> DNA <213> human SCK / SLI gene minisatellite <400> 15 atctgcgtgc tcgtttgcgt gctc 24 <210> 16 <211> 24 <212> DNA <213> human SCK / SLI gene minisatellite <400> 16 atctgcgtgc tcgtctgcgt gctc 24 <210> 17 <211> 24 <212> DNA <213> human SCK / SLI gene minisatellite <400> 17 atctgcatgc tcatttgcat gctc 24 <210> 18 <211> 24 <212> DNA <213> human SCK / SLI gene minisatellite <400> 18 atctgcatgc ttgtttgcgt gctc 24 <210> 19 <211> 38 <212> DNA <213> human SCK / SLI gene minisatellite <400> 19 gtcatatttc agcgtgtgga tggccacgcc gtggccac 38 <210> 20 <211> 38 <212> DNA <213> human SCK / SLI gene minisatellite <400> 20 gtcatatttc agcgtgtgga tggccacgcc gtggccac 38 <210> 21 <211> 38 <212> DNA <213> human SCK / SLI gene minisatellite <400> 21 gtcgtatttc agcgtgtgga tggccacgcc atggccac 38 <210> 22 <211> 38 <212> DNA <213> human SCK / SLI gene minisatellite <400> 22 accatatttc agcgtgtgga tggccacgcc atggccac 38 <210> 23 <211> 65 <212> DNA <213> human SCK / SLI gene minisatellite <400> 23 tgcgtttctc cgtttcattg aggttggggc atcgtactga cttgggggga attcctgcgt 60 aggtg 65 <210> 24 <211> 64 <212> DNA <213> human SCK / SLI gene minisatellite <400> 24 tgcgtttctc cgtttcattg aggttggggc atcgtactga cttgggggag ttcctgcgta 60 ggtg 64 <210> 25 <211> 64 <212> DNA <213> human SCK / SLI gene minisatellite <400> 25 tgtgtttctc cgtttcattg aggttggggc atcgtactga cttgggggag ttcctgcgta 60 ggtg 64 <210> 26 <211> 63 <212> DNA <213> human SCK / SLI gene minisatellite <400> 26 tgcacctgct gtgccccgcc tagaactctg tctgcaaacc ccaccacgtc aggcctttgc 60 acc 63 <210> 27 <211> 62 <212> DNA <213> human SCK / SLI gene minisatellite <400> 27 tgcacctgct gtgccccgcc tagaactctg tctgcaaacc caccacgtca ggcctttgca 60 cc 62 <210> 28 <211> 62 <212> DNA <213> human SCK / SLI gene minisatellite <400> 28 tgcacctgct gtccccccct agaactctgt ctgcaaactc caccacgtca ggcctttgca 60 cc 62 <210> 29 <211> 63 <212> DNA <213> human SCK / SLI gene minisatellite <400> 29 tgcacctgct gtgccccgcc tagaactctg tctgcaaact ccaccgcatc gggcctttcc 60 acc 63 <210> 30 <211> 21 <212> DNA <213> human SCK / SLI gene minisatellite <400> 30 ggggaggcgg aagcgggtcc c 21 <210> 31 <211> 21 <212> DNA <213> human SCK / SLI gene minisatellite <400> 31 ggggaggcgg aagcgggtcc c 21 <210> 32 <211> 21 <212> DNA <213> human SCK / SLI gene minisatellite <400> 32 ggggaggcgg aagcgggtcc t 21 <210> 33 <211> 29 <212> DNA <213> human SCK / SLI gene minisatellite <400> 33 gtagggggaa ggaccccact gaacccagc 29 <210> 34 <211> 29 <212> DNA <213> human SCK / SLI gene minisatellite <400> 34 gtagggggaa ggaccccact gaactcagt 29 <210> 35 <211> 29 <212> DNA <213> human SCK / SLI gene minisatellite <400> 35 gtagggggaa ggaccccact gaacccagc 29 <210> 36 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR1_forwar primer <400> 36 ggcggccttg aattcctcca 20 <210> 37 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> TR1_reverse primer <400> 37 ctcagccccc agggtcagg 19 <210> 38 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> TR1_forward primer <400> 38 tgacaggcca ctctaaggct ctcc 24 <210> 39 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> TR1_reverse primer <400> 39 atgctgactg gccaggctag gagtt 25 <210> 40 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> TR2_forwar primer <400> 40 ggtcaggatg ggtgagaggg aag 23 <210> 41 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR2_reverse primer <400> 41 cctgcgtctc gcactgtcgt 20 <210> 42 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR3_forwar primer <400> 42 acctgaggga gcgggaagga 20 <210> 43 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR3_reverse primer <400> 43 tgccatgtgg cctccctttt 20 <210> 44 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> TR3_forward primer <400> 44 gatctcaatg cccatcagga atg 23 <210> 45 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> TR3_reverse primer <400> 45 ctgggctcaa gagatcctcc cttc 24 <210> 46 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> TR4_forwar primer <400> 46 ccatgatgtt ttgaaccatg tgga 24 <210> 47 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> TR4_reverse primer <400> 47 agatcgtctt gctcctaaca ctctgc 26 <210> 48 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> TR4_forwar primer <400> 48 gggaggcaaa gaaaacagat gcag 24 <210> 49 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR7_forward primer <400> 49 gctgcaaggg gtacccacga 20 <210> 50 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> TR7_forwar primer <400> 50 gaagtcatgt ctgaaatgcc acctc 25 <210> 51 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR7_reverse primer <400> 51 cacaatgggg cgccgtgtat 20 <210> 52 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR8_forward primer <400> 52 ttctttcctt ggccgcatca 20 <210> 53 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR8_reverse primer <400> 53 ccagcagccc aaaggtggaa 20 <210> 54 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TR9_forward primer <400> 54 caggaatggt ggcctttgtg g 21 <210> 55 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TR9_reverse primer <400> 55 cccacctacg caggaactcc a 21 <210> 56 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> TR9_forward primer <400> 56 gggaggcaaa gaaaacagat gcag 24 <210> 57 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR9_reverse primer <400> 57 gctgcaaggg gtacccacga 20 <210> 58 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> TR10 forward primer <400> 58 cccgctccct ctccatcag 19 <210> 59 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR10_reverse primer <400> 59 gccgtggggt ttgcagacat 20 <210> 60 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR10 forward primer <400> 60 acactctgcg ccacccccta 20 <210> 61 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR10_reverse primer <400> 61 ctcccaggag gcgaccttga 20 <210> 62 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR11_forward primer <400> 62 aggacaaacc gccgggagac 20 <210> 63 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> TR11_reverse primer <400> 63 agcccccaga agctggct 18 <210> 64 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TR12_forward primer <400> 64 tgcaaggacc ctgttcccaa t 21 <210> 65 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR12_reverse primer <400> 65 ggtgggtgct cccagctcct 20 <210> 66 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> TR12_forward primer <400> 66 agagacccct gatccaggta ggc 23 <210> 67 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR12_reverse primer <400> 67 tgcggccagg aacattagcc 20 <210> 68 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR12_forward primer <400> 68 acagccagct tctgggggct 20 <210> 69 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> TR12_reverse primer <400> 69 tgcggccagg aacattagcc 20

Claims (13)

A mini satellite marker composition for identifying a primate or primate individual, comprising at least one mini satellite represented by any one or more of the nucleotide sequences of SEQ ID NOS: 1 to 35.
2. The miniature satellite marker composition according to claim 1, wherein the primate is at least one selected from the group consisting of a human, a chimpanzee, a gorilla, an orangutan, and a rhesus monkey.
1, 4, 7, 10, 15, 19, 23, 26, 30, and 33 are miniature satellites for identifying humans; SEQ ID NOS: 2, 5, 8, 11, 16, 20, 24, 27, 31 and 34 are mini satellite for identifying chimpanzees; SEQ ID NOS: 3, 6, 9, 12, 17, 21, 25, 28, 32 and 35 are mini satellites for identifying gorillas; SEQ ID NOS: 13, 18, 22 and 29 are mini-satellite to identify orangutans; And SEQ ID NO: 14 is mini satellite for identifying a Rhesus monkey; and a mini satellite marker composition for identifying primate or primate individuals.
SEQ ID NO: 36 and SEQ ID NO: 37; SEQ ID NO: 38 and SEQ ID NO: 39; SEQ ID NO: 40 and SEQ ID NO: 41; SEQ ID NO: 42 and SEQ ID NO: 43; SEQ ID NO: 44 and SEQ ID NO: 45; SEQ ID NO: 46 and SEQ ID NO: 47; SEQ ID NO: 48 and SEQ ID NO: 49; SEQ ID NO: 50 and SEQ ID NO: 51; SEQ ID NO: 52 and SEQ ID NO: 53; SEQ ID NO: 54 and SEQ ID NO: 55; SEQ ID NO: 56 and SEQ ID NO: 57; SEQ ID NO: 58 and SEQ ID NO: 59; SEQ ID NO: 60 and SEQ ID NO: 61; SEQ ID NO: 62 and SEQ ID NO: 63; SEQ ID NO: 64 and SEQ ID NO: 65; SEQ ID NO: 66 and SEQ ID NO: 67; And a primer set for amplifying a miniature satellite for identifying a primate or primate object comprising at least one primer pair selected from the group consisting of SEQ ID NO: 68 and SEQ ID NO: 69.
5. The method according to claim 4, wherein the primer set
SEQ ID NO: 36 and SEQ ID NO: 37; SEQ ID NO: 40 and SEQ ID NO: 41; SEQ ID NO: 42 and SEQ ID NO: 43; SEQ ID NO: 46 and SEQ ID NO: 47; SEQ ID NO: 50 and SEQ ID NO: 51; SEQ ID NO: 52 and SEQ ID NO: 53; SEQ ID NO: 54 and SEQ ID NO: 55; SEQ ID NO: 58 and SEQ ID NO: 59; SEQ ID NO: 62 and SEQ ID NO: 63; And a primer set for mini satellite amplification for identifying a human comprising at least one primer pair selected from the group consisting of SEQ ID NO: 64 and SEQ ID NO: 65;
SEQ ID NO: 38 and SEQ ID NO: 39; SEQ ID NO: 40 and SEQ ID NO: 41; SEQ ID NO: 44 and SEQ ID NO: 45; SEQ ID NO: 46 and SEQ ID NO: 47; SEQ ID NO: 50 and SEQ ID NO: 51; SEQ ID NO: 52 and SEQ ID NO: 53; SEQ ID NO: 56 and SEQ ID NO: 57; SEQ ID NO: 58 and SEQ ID NO: 59; SEQ ID NO: 62 and SEQ ID NO: 63; SEQ ID NO: 66 and SEQ ID NO: 67; A primer set for mini satellite amplification for identifying a chimpanzee comprising at least one primer pair selected from the group consisting of SEQ ID NO: 68 and SEQ ID NO: 69;
SEQ ID NO: 38 and SEQ ID NO: 39; SEQ ID NO: 40 and SEQ ID NO: 41; SEQ ID NO: 44 and SEQ ID NO: 45; SEQ ID NO: 46 and SEQ ID NO: 47; SEQ ID NO: 50 and SEQ ID NO: 51; SEQ ID NO: 52 and SEQ ID NO: 53; SEQ ID NO: 56 and SEQ ID NO: 57; SEQ ID NO: 60 and SEQ ID NO: 61; SEQ ID NO: 62 and SEQ ID NO: 63; SEQ ID NO: 66 and SEQ ID NO: 67; A primer set for mini satellite amplification for identifying a gorilla comprising at least one primer pair selected from the group consisting of SEQ ID NO: 68 and SEQ ID NO: 69;
SEQ ID NO: 48 and SEQ ID NO: 49; SEQ ID NO: 50 and SEQ ID NO: 51; SEQ ID NO: 52 and SEQ ID NO: 53; And a primer set for mini satellite amplification for identifying orangutans comprising at least one primer pair selected from the group consisting of SEQ ID NO: 58 and SEQ ID NO: 59;
SEQ ID NO: 46 and SEQ ID NO: 47; and a primer set for mini satellite amplification for identifying a resus monkey comprising a pair of primers consisting of a primer pair consisting of SEQ ID NO: set.
5. The primer set according to claim 4, wherein the primate is at least one selected from the group consisting of a human, a chimpanzee, a gorilla, an orangutan, and a rhesus monkey.
A PCR kit for identifying primate or primate individuals, comprising the primer set of claim 4.
A probe for identifying a primate or a primate, comprising at least one base sequence selected from SEQ ID NOS: 1 to 35, or a nucleotide represented by its complementary base sequence.
delete A microarray for primate or primate identification, comprising a nucleotide represented by any one or more of the nucleotide sequences selected from SEQ ID NOS: 1 to 35, a polypeptide encoded by the polynucleotide, or cDNA thereof.
1) SEQ ID NO: 36 and SEQ ID NO: 37; SEQ ID NO: 38 and SEQ ID NO: 39; SEQ ID NO: 40 and SEQ ID NO: 41; SEQ ID NO: 42 and SEQ ID NO: 43; SEQ ID NO: 44 and SEQ ID NO: 45; SEQ ID NO: 46 and SEQ ID NO: 47; SEQ ID NO: 48 and SEQ ID NO: 49; SEQ ID NO: 50 and SEQ ID NO: 51; SEQ ID NO: 52 and SEQ ID NO: 53; SEQ ID NO: 54 and SEQ ID NO: 55; SEQ ID NO: 56 and SEQ ID NO: 57; SEQ ID NO: 58 and SEQ ID NO: 59; SEQ ID NO: 60 and SEQ ID NO: 61; SEQ ID NO: 62 and SEQ ID NO: 63; SEQ ID NO: 64 and SEQ ID NO: 65; SEQ ID NO: 66 and SEQ ID NO: 67; A primer set comprising at least one primer pair selected from the group consisting of SEQ ID NO: 68 and SEQ ID NO: 69, and a whole DNA sample extracted from the primate;
2) DNA sequencing using the amplified DNA sample obtained in step 1); And
3) analyzing the DNA sequencing result of step 2) in comparison with a miniature satellite represented by any one or more of the nucleotide sequences selected from SEQ ID NOS: 1 to 35, and identifying the primate or primate.
12. The primate or primate identification method according to claim 11, wherein the primate in step 1) is at least one selected from the group consisting of a human, a chimpanzee, a gorilla, an orangutan, and a Rhesus monkey.
12. The method according to claim 11, wherein the primer set in step 1)
SEQ ID NO: 36 and SEQ ID NO: 37; SEQ ID NO: 40 and SEQ ID NO: 41; SEQ ID NO: 42 and SEQ ID NO: 43; SEQ ID NO: 46 and SEQ ID NO: 47; SEQ ID NO: 50 and SEQ ID NO: 51; SEQ ID NO: 52 and SEQ ID NO: 53; SEQ ID NO: 54 and SEQ ID NO: 55; SEQ ID NO: 58 and SEQ ID NO: 59; SEQ ID NO: 62 and SEQ ID NO: 63; And a primer set for mini satellite amplification for identifying a human comprising at least one primer pair selected from the group consisting of SEQ ID NO: 64 and SEQ ID NO: 65;
SEQ ID NO: 38 and SEQ ID NO: 39; SEQ ID NO: 40 and SEQ ID NO: 41; SEQ ID NO: 44 and SEQ ID NO: 45; SEQ ID NO: 46 and SEQ ID NO: 47; SEQ ID NO: 50 and SEQ ID NO: 51; SEQ ID NO: 52 and SEQ ID NO: 53; SEQ ID NO: 56 and SEQ ID NO: 57; SEQ ID NO: 58 and SEQ ID NO: 59; SEQ ID NO: 62 and SEQ ID NO: 63; SEQ ID NO: 66 and SEQ ID NO: 67; A primer set for mini satellite amplification for identifying a chimpanzee comprising at least one primer pair selected from the group consisting of SEQ ID NO: 68 and SEQ ID NO: 69;
SEQ ID NO: 38 and SEQ ID NO: 39; SEQ ID NO: 40 and SEQ ID NO: 41; SEQ ID NO: 44 and SEQ ID NO: 45; SEQ ID NO: 46 and SEQ ID NO: 47; SEQ ID NO: 50 and SEQ ID NO: 51; SEQ ID NO: 52 and SEQ ID NO: 53; SEQ ID NO: 56 and SEQ ID NO: 57; SEQ ID NO: 60 and SEQ ID NO: 61; SEQ ID NO: 62 and SEQ ID NO: 63; SEQ ID NO: 66 and SEQ ID NO: 67; A primer set for mini satellite amplification for identifying a gorilla comprising at least one primer pair selected from the group consisting of SEQ ID NO: 68 and SEQ ID NO: 69;
SEQ ID NO: 48 and SEQ ID NO: 49; SEQ ID NO: 50 and SEQ ID NO: 51; SEQ ID NO: 52 and SEQ ID NO: 53; And a primer set for mini satellite amplification for identifying orangutans comprising at least one primer pair selected from the group consisting of SEQ ID NO: 58 and SEQ ID NO: 59;
SEQ ID NO: 46 and SEQ ID NO: 47; and a primer set for mini satellite amplification for identifying a resus monkey comprising a pair of primers consisting of SEQ ID NO: 46 and SEQ ID NO: 47.

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