CN1519367A - Polymorphism of 31th exon in PCNT gene - Google Patents
Polymorphism of 31th exon in PCNT gene Download PDFInfo
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- CN1519367A CN1519367A CNA031150624A CN03115062A CN1519367A CN 1519367 A CN1519367 A CN 1519367A CN A031150624 A CNA031150624 A CN A031150624A CN 03115062 A CN03115062 A CN 03115062A CN 1519367 A CN1519367 A CN 1519367A
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Abstract
A single nucleotide polymorphism (SNP) for the extron No.31 of PCNT gene and a method for analyzing the SNP and activity of said extron No.31 are disclosed. Said SNP is T->C polymorphism at the site No.250 in its sequence shown by SEQ ID No.1 and can cause the change Leu(L)->Pro(P) at the site No.83 of coding protein, so changing protein activity.
Description
Technical field
The present invention relates to the single nucleotide polymorphism of PCNT gene hentriaconta-exon.The invention still further relates to the method for the allelic mutation of analyzing PCNT gene hentriaconta-exon.
Background technology
(pericentrin PCNT) is a kind of biomass cells centrosome associated protein of high conservative to middle peripheral proteins.The investigator thinks, it is closely related with the forming process of centrosome microtubule tissue in the eukaryotic cell.(Cell?1994Feb?25；76(4)：639-50)。
Before the application, also there is not the relevant report of PCNT gene hentriaconta-exon polymorphism.
Summary of the invention
Purpose of the present invention just provides the polymorphism and the detection method thereof of PCNT gene hentriaconta-exon.
In a first aspect of the present invention, a kind of method of single nucleotide polymorphism of the people of detection PCNT gene hentriaconta-exon is provided, it comprises step:
(a) determine the 250th Nucleotide in sequence shown in the SEQ ID NO:1 of people PCNT gene hentriaconta-exon;
(b) detect whether there is single nucleotide polymorphism in described position.
In a preference, described single nucleotide polymorphism is to have C at the 250th.
In a second aspect of the present invention, a kind of isolating nucleic acid is provided, it has the sequence shown in the SEQ ID NO:1, and the 250th is C.
In a third aspect of the present invention, a kind of allele specific nucleic acid primer is provided, its length is 15-50bp, and hybridizes specifically and amplify shown in the SEQ IDNO:1 that contains people PCNT gene hentriaconta-exon the amplified production of the 250th single nucleotide polymorphism in the sequence.
In a fourth aspect of the present invention, a kind of allele specific oligonucleotide probe is provided, its length is 15-50bp, and hybridizes specifically and detect shown in the SEQ IDNO:1 that contains people PCNT gene hentriaconta-exon the 250th single nucleotide polymorphism in the sequence.
A kind of test kit comprises:
(1) primer of specific amplification people PCNT gene hentriaconta-exon is right,
(2) detect amplified production and compare the reagent that whether exists variation required, the 250th single nucleotide polymorphism in the sequence shown in the described SEQ ID NO:1 with normal PCNT gene hentriaconta-exon.
Another kind of diagnostic kit comprises: the oligonucleotide probe that the primer that the present invention is above-mentioned and/or the present invention are above-mentioned.
Embodiment
The inventor is by extensive and deep research, have been found that in the coded protein sequence of PCNT gene hentriaconta-exon the 83rd Leu (L)-Pro (P) polymorphic (be last 250 T-of mRNA〉C polymorphism), finished the present invention on this basis.
According to normal people PCNT gene sequencing result, found this SNP.Find that by analysis this SNP causes the 83rd amino acids by Leu (L)-Pro (P).Because this is the amino acid whose displacement of different properties, therefore, can cause the coded protein-active of PCNT gene hentriaconta-exon to change.
The cDNA sequence of PCNT gene hentriaconta-exon is listed in SEQ ID NO:1, and wherein open reading frame is the 3-350 position, and its coded aminoacid sequence is listed in SEQ ID NO:2.The genome sequence of PCNT gene hentriaconta-exon can obtain from GenBank.
Those skilled in the art will appreciate that a large amount of analytical technologies can be used for detecting whether the site exists single nucleotide polymorphism described in the PCNT gene hentriaconta-exon.These technology comprise (but being not limited to): dna sequencing, sequencing by hybridization; Enzymatic mispairing cutting, heteroduple analysis, dot blot, oligonucleotide arrays (DNA chip), Mini-sequencing, Taqman technology, molecular beacon etc., sex change high performance liquid chromatography (DHPLC).
On the other hand, detection method of the present invention is used to assess the susceptibility of the individual PCNT of suffering from gene hentriaconta-exon relative disease.
Being used for specimen of the present invention and being not particularly limited, for detecting SNP, can be DNA or the mRNA that extracting goes out from samples such as blood, tissue.For detect PCNT gene hentriaconta-exon active for, can anyly contain the sample of PCNT gene hentriaconta-exon, as blood etc.
The primer and the probe that are used for the inventive method or detection kit design according to the cDNA sequence (sequence of SEQ ID NO:1) or the genome sequence of PCNT gene hentriaconta-exon, and the synthetic technology of available routine is synthesized and got final product.
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in and limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, people such as Sambrook for example, molecular cloning: laboratory manual (New York:ColdSpring Harbor Laboratory Press, 1989) condition described in, or the condition of advising according to manufacturer.
Embodiment 1
The extracting of gene and order-checking
Extract DNA with conventional phenol chloroform method from people's blood, concentration correction is used for conventional pcr amplification to 20ng/ul.Primer is:
Adopted primer is arranged: 5 '-cttcaccgac cagcgtactt-3 ' (SEQ ID NO:3)
Antisense primer: 5 '-gtgtgctgtc aggaggtgaa-3 ' (SEQ ID NO:4)
The product of the 451bp that amplifies carries out dna sequencing behind the purifying.
Embodiment 2
The acquisition of SNP
The PCNT gene is directly checked order.Each sample sequence of measuring is compared, thereby draw the difference of sequence, obtain SNP.
Embodiment 3
Detection kit
Preparation one detects the detection kit of PCNT gene-correlation disease susceptibility, and wherein, it is right to contain the following primer that amplifies 250 SNP:
Adopted primer is arranged: 5 '-cttcaccgac cagcgtactt-3 ' (SEQ ID NO:3)
Antisense primer: 5 '-gtgtgctgtc aggaggtgaa-3 ' (SEQ ID NO:4)
Amplified production and normal control are carried out stratographic analysis with sex change high performance liquid chromatograph (DHPLC), can detect 250 T-easily C type SNP.
All quote in this application as a reference at all documents that the present invention mentions, just quoted as a reference separately as each piece document.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after having read above-mentioned teachings of the present invention, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
SEQUENCE?LISTING
＜110〉Research Center of Shanghai Human Genome
＜120〉PCNT gene hentriaconta-exon polymorphism
<130>030115
<160>4
<170>PatentIn?version?3.1
<210>1
<211>451
<212>DNA
<213>Homo?sapiens
<220>
<221>CDS
<222>(3)..(350)
<223>
<400>1
ct?tca?ccg?acc?agc?gta?ctt?ggt?ggc?tcc?cgc?cac?cag?agc?cac?act 47
Ser?Pro?Thr?Ser?Val?Leu?Gly?Gly?Ser?Arg?His?Gln?Ser?His?Thr
1 5 10 15
gca?gag?gct?ggg?ccc?cgg?aag?agc?ccg?gtc?ggg?atg?ctg?gac?ctg?tct 95
Ala?Glu?Ala?Gly?Pro?Arg?Lys?Ser?Pro?Val?Gly?Met?Leu?Asp?Leu?Ser
20 25 30
tcc?tgg?agc?tcc?cct?gag?gtc?ctc?agg?aag?gac?tgg?acc?ctg?gag?ccc 143
Ser?Trp?Ser?Ser?Pro?Glu?Val?Leu?Arg?Lys?Asp?Trp?Thr?Leu?Glu?Pro
35 40 45
tgg?ccc?agc?ctc?ccc?gtg?aca?ccc?cac?tca?gga?gcc?ctg?agc?ctg?tgc 191
Trp?Pro?Ser?Leu?Pro?Val?Thr?Pro?His?Ser?Gly?Ala?Leu?Ser?Leu?Cys
50 55 60
agt?gcc?gac?aca?tcc?ctg?ggg?gac?agg?gcg?gac?acc?tcg?ctg?cca?cag 239
Ser?Ala?Asp?Thr?Ser?Leu?Gly?Asp?Arg?Ala?Asp?Thr?Ser?Leu?Pro?Gln
65 70 75
acc?cag?ggg?ctg?ggg?ctg?ctt?tgt?tcc?cca?ggc?gtg?tct?gca?gca?gcg 287
Thr?Gln?Gly?Leu?Gly?Leu?Leu?Cys?Ser?Pro?Gly?Val?Ser?Ala?Ala?Ala
80 85 90 95
ctg?gca?ctg?cag?tgg?gcc?gag?tct?ccg?ccg?gct?gac?gac?cac?cat?gtg 335
Leu?Ala?Leu?Gln?Trp?Ala?Glu?Ser?Pro?Pro?Ala?Asp?Asp?His?His?Val
100 105 110
cag?agg?acg?gct?gtg?gtaggtgcct?gctctgctcc?caggcctgct?gttcccgtgg 390
Gln?Arg?Thr?Ala?Val
115
gaatgtggtc?tgcccggcgc?cacggcagct?gccattgttt?gttcacctcc?tgacagcaca 450
c 451
<210>2
<211>116
<212>PRT
<213>Homo?sapiens
<400>2
Ser?Pro?Thr?Ser?Val?Leu?Gly?Gly?Ser?Arg?His?Gln?Ser?His?Thr?Ala
1 5 10 15
Glu?Ala?Gly?Pro?Arg?Lys?Ser?Pro?Val?Gly?Met?Leu?Asp?Leu?Ser?Ser
20 25 30
Trp?Ser?Ser?Pro?Glu?Val?Leu?Arg?Lys?Asp?Trp?Thr?Leu?Glu?Pro?Trp
35 40 45
Pro?Ser?Leu?Pro?Val?Thr?Pro?His?Ser?Gly?Ala?Leu?Ser?Leu?Cys?Ser
50 55 60
Ala?Asp?Thr?Ser?Leu?Gly?Asp?Arg?Ala?Asp?Thr?Ser?Leu?Pro?Gln?Thr
65 70 75 80
Gln?Gly?Leu?Gly?Leu?Leu?Cys?Ser?Pro?Gly?Val?Ser?Ala?Ala?Ala?Leu
85 90 95
Ala?Leu?Gln?Trp?Ala?Glu?Ser?Pro?Pro?Ala?Asp?Asp?His?His?Val?Gln
100 105 110
Arg?Thr?Ala?Val
115
<210>3
<211>20
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>3
cttcaccgac?cagcgtactt 20
<210>4
<211>20
<212>DNA
＜213〉artificial sequence
<220>
<221>misc_feature
＜223〉primer
<400>4
gtgtgctgtc?aggaggtgaa 20
Claims (7)
1. method that detects the single nucleotide polymorphism of people PCNT gene hentriaconta-exon is characterized in that it comprises step:
(a) determine the 250th Nucleotide in sequence shown in the SEQ ID NO:1 of people PCNT gene hentriaconta-exon;
(b) detect whether there is single nucleotide polymorphism in described position.
2. the method for claim 1 is characterized in that, described single nucleotide polymorphism is to have C at the 250th.
3. an isolating nucleic acid is characterized in that, it has the sequence shown in the SEQ ID NO:1, and the 250th is C.
4. allele specific nucleic acid primer, it is characterized in that, its length is 15-50bp, and hybridizes specifically and amplify shown in the SEQ ID NO:1 that contains people PCNT gene hentriaconta-exon the amplified production of the 250th single nucleotide polymorphism in the sequence.
5. an allele specific oligonucleotide probe is characterized in that, its length is 15-50bp, and hybridizes specifically and detect shown in the SEQ ID NO:1 that contains people PCNT gene hentriaconta-exon the 250th single nucleotide polymorphism in the sequence.
6. diagnostic kit is characterized in that it comprises:
(1) primer of specific amplification people PCNT gene hentriaconta-exon is right,
(2) detect amplified production and compare the reagent that whether exists variation required, the 250th single nucleotide polymorphism in the sequence shown in the described SEQ ID NO:1 with normal PCNT gene hentriaconta-exon.
7. a diagnostic kit is characterized in that it comprises: described primer of claim 4 and/or the described oligonucleotide probe of claim 5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA031150624A CN1519367A (en) | 2003-01-22 | 2003-01-22 | Polymorphism of 31th exon in PCNT gene |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA031150624A CN1519367A (en) | 2003-01-22 | 2003-01-22 | Polymorphism of 31th exon in PCNT gene |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1519367A true CN1519367A (en) | 2004-08-11 |
Family
ID=34284106
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA031150624A Pending CN1519367A (en) | 2003-01-22 | 2003-01-22 | Polymorphism of 31th exon in PCNT gene |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1519367A (en) |
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2003
- 2003-01-22 CN CNA031150624A patent/CN1519367A/en active Pending
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