CN111172169A - Non-syndrome type congenital missing tooth related low-frequency/rare mutation and detection method thereof - Google Patents
Non-syndrome type congenital missing tooth related low-frequency/rare mutation and detection method thereof Download PDFInfo
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Abstract
The invention discloses a low-frequency/rare mutation related to non-syndrome type congenital missing tooth and a detection method thereof. The FGFR1 gene is mutated into a single point mutation c.G103A (chr 8: 38287455) on a third exon, and the amino acid is changed into p.G35R. The pathogenic gene FGFR1 provided by the invention can provide a basis for screening and detecting the pathogenic gene and making a treatment scheme and the like. Meanwhile, the invention constructs a pathogenic gene detection method; the non-syndromic congenital missing tooth pedigree was collected as a study model, whole genome exon sequencing was used, bioinformatics analysis was performed, and Sanger sequencing was followed to verify mutations and annotate related genes.
Description
Technical Field
The invention belongs to the field of medical biomedicine, and relates to a low-frequency/rare mutation related to non-syndrome type congenital missing tooth and a detection method thereof.
Background
Congenital missing teeth (Tooth-deficiency) refers to a congenital decrease in the number of teeth due to abnormal development of Tooth germ. Non-syndromic tooth loss (non-syndromic tooth loss) and syndromic tooth loss (syndromic tooth loss) can be classified according to whether systemic symptoms are accompanied. The incidence of congenital tooth loss is reported to be 2.2% to 10.1% (excluding the third molar loss) and women have a higher incidence than men.
Congenital missing teeth (Tooth administration) classification: according to the number of the missing teeth, the missing teeth can be divided into a few missing teeth (the number of the missing teeth is less than 6, most common missing teeth of a third molar), a plurality of missing teeth (the number of the missing teeth is more than or equal to 6 except the third molar) and congenital missing teeth (all the congenital missing teeth). The non-syndromic congenital deficient dental caries is considered to be genetically related, and its genetic mode involves autosomal dominant inheritance, autosomal recessive inheritance and X-linked inheritance, which can be sporadic or familial.
The discovery of pathogenic genes and mutations related to the congenital missing tooth has important significance for the diagnosis and screening of the congenital missing tooth.
Disclosure of Invention
The invention provides a low-frequency/rare mutation related to non-syndrome type congenital missing tooth, and simultaneously, a non-syndrome type congenital missing tooth family is used as a research model of diseases related to non-syndrome type congenital missing tooth, and a basis can be provided for analysis of a non-syndrome type congenital missing tooth pathogenesis, pathogenic gene screening and detection, formulation of a treatment scheme and the like by combining a whole genome exon sequencing and Sanger sequencing detection method.
The invention provides a low-frequency/rare mutation related to non-syndrome type congenital missing tooth, wherein a mutant gene is FGFR1, and the sequence after the mutation is shown in SEQ ID NO. 3.
Compared with a wild FGFR1 gene, the 103 th base on the third exon of the FGFR1 gene sequence is mutated from G to A in the FGFR1 mutant gene related to non-syndrome type congenital missing teeth.
The FGFR1 gene mutation related in the invention is detected and verified to accord with autosomal dominant inheritance, is a non-syndrome congenital tooth missing pathogenic gene, and is a heterozygous mutation FGFR1: exon3: c.G103A. The transcriptome number of the wild FGFR1 gene in NCBI database is NM-001174063, the 103 th base of the gene in the third exon is mutated from G to A, and other parts are the same as the wild type. Since the mutation occurs in the exon region, SEQ ID NO.4 provided by the present invention is the gene sequence of FGFR1 wild type for reference.
The invention also provides a mutant FGFR1 protein, NP-001167534 and p.G35R, wherein the 35 th amino acid of the wild-type protein is mutated from glycine to arginine, namely the protein is obtained by mutating the 35 th amino acid of the wild-type FGFR1 protein from glycine to arginine, and other parts of the protein are the same as the wild type.
The wild-type FGFR1 protein is numbered in the NCBI database as: NP _ 001167534.
The invention also provides application of the FGFR1 mutant gene in non-syndrome type congenital tooth missing mechanism research, screening, auxiliary diagnosis and/or treatment.
The invention also provides a detection method of the FGFR1 mutant gene, which comprises the following main steps:
(1) extracting genome DNA;
(2) sanger sequencing: and (3) carrying out genotype detection on the FGFR1 mutant gene synthesis primer, wherein the 103 th base in the third exon of the FGFR1 gene sequence is mutated from G to A and is an indication of non-syndrome type congenital missing teeth.
In the detection method of the present invention, the genomic DNA extracted in step (1) may be a biological sample commonly used in the art, such as blood.
In one embodiment, in the detection method of the present invention, the primer sequence in step (2) is as shown in SEQ ID NO: 1 and SEQ ID NO: 2, respectively.
The invention also provides a primer for detecting the FGFR1 mutant gene, which has a sequence shown in SEQ ID NO: 1 and SEQ ID NO: 2, respectively.
The invention also provides application of the primer in preparation of a reagent for screening FGFR1 mutant genes.
A kit for screening FGFR1 mutant genes comprises the primers provided by the invention, and the kit can further comprise a DNA extraction reagent.
The invention is based on a genetic family, combines a whole genome exon sequencing and Sanger sequencing detection method, and comprises the following main steps:
(1) family collection and standardized oral examination of family members: intraoral congenital missing teeth (excluding third molar missing), inquiring whether family missing genetic history exists (family history without congenital missing teeth), signing informed consent, and collecting family member peripheral blood;
(2) auxiliary inspection: panoramic film: mother 13, 23 missing (except for the third molar loss), son 15, 13, 23, 25, 35, 33, 31, 41, 43, 45 missing (except for the third molar loss), father without missing teeth;
(3) gene detection: extracting peripheral blood, extracting DNA, and screening out pathogenic genes by sequencing exon of whole genome;
(4) screening candidate genes: bioinformatics analysis was performed according to 1000 Gemomes Project, ESP6500siv2-ALL, genomAD-ALL and genomAD-EAS and SIFT, PolyPhen-2, Mutation Taster etc. software and followed genotype-phenotype co-segregation.
Sanger sequencing verification: verification is carried out in all family members to determine whether the family members carry the gene and simultaneously accord with an autosomal dominant inheritance pattern.
In one embodiment of the present invention, a more specific method for detecting a pathogenic gene of a non-syndromic missing tooth is further provided, which comprises the following main steps:
(1) a detailed family history survey (family history of no congenital missing teeth) was performed on the patient; a normative oral examination was performed, patient signs: intraoral tooth loss (excluding third molar loss); signing an informed consent, collecting peripheral blood of family members, and extracting DNA;
(2) capturing and enriching DNA of a whole genome exon region by using a sequence capture technology, and then performing high-throughput sequencing on an Illumina platform;
(3) performing bioinformatics analysis on the sequencing result, and screening out candidate mutant genes according to genotype phenotype cosegregation analysis;
(4) sanger sequencing is carried out on a patient family related member sample, the sequence of a PCR forward primer is shown as SEQ ID No.1, and the sequence of a PCR reverse primer is shown as SEQ ID No. 2.
The invention has the advantages that: the non-syndromic congenital missing tooth is a clinically common missing tooth type, and the influence of the FGFR1 gene on tooth development is discovered through gene detection of patients of the genetic family. Therefore, the family can be used as a research model of the diseases related to the non-syndrome type congenital missing tooth and has potential important values for clinical gene diagnosis, screening, accurate treatment and the like of the diseases related to the non-syndrome type congenital missing tooth and the like. The mutant gene can provide basis and lay the foundation for the analysis of the pathogenesis of non-syndrome type congenital missing tooth, the development of medicines, the screening and detection of the pathogenesis gene, the formulation of a treatment scheme and the like.
Drawings
FIG. 1 is a pedigree D map of a non-syndromic congenital missing tooth;
FIG. 2 is a panorama of all members of a family;
FIG. 3 is a flowchart for screening a pathogenic gene;
FIG. 4 is a Sanger sequencing validation result in which the wild type was the result of genotyping I1 and the mutant was the result of genotyping I2 and II 1.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. It is intended that all modifications or alterations to the methods, procedures or conditions of the present invention be made without departing from the spirit or essential characteristics thereof.
Example 1
1. Selection of samples (all Chinese Han nationality)
The inventor collects a non-syndromic congenital tooth-missing family with definite diagnosis from the oral hospital affiliated to the university of medical Nanjing. Three individuals in pedigree D (fig. 1) were examined and found to have a deletion in mothers 13, 23 (with the exception of the third molar deletion), loss in children 15, 13, 23, 25, 35, 33, 31, 41, 43, 45 (with the exception of the third molar deletion), and a normal paternal phenotype (fig. 2). Everyone in the family is not accompanied by other related systemic diseases. In the process of diagnosing and treating the genetic diseases, patients have better compliance and subjective treatment willingness, all study subjects sign informed consent (under 18 years old, signed by guardians of the study subjects), complete corresponding epidemiological investigation through ethical approval of medical institutions, and provide 1-3mL of peripheral blood specimens.
2. Genomic DNA extraction
Extracting the genome DNA of each research object by using a QIAGEN kit, precipitating by using isopropanol to obtain the genome DNA, dissolving the DNA precipitate by AE to determine the concentration, diluting, subpackaging and storing in a refrigerator at the temperature of-20 ℃ for later use.
The method comprises the following specific steps:
(1) adding 500 mul of cell lysate CL into 400 mul of anticoagulation blood, and reversing and mixing uniformly for 5 times;
(2) centrifuging at 12,000rpm (13,400 Xg) for 1min, and discarding the supernatant;
(3) repeating the steps 1 and 2 to fully lyse the cells;
(4) adding 200 mul of buffer solution FG and Protease K mixed solution, immediately whirling and uniformly mixing until the solution has no lumps;
(5) water bath at 65 deg.C for 15min, and mixing by reversing for several times;
(6) adding 200 mul of isopropanol, reversing, fully and uniformly mixing until filamentous or clustered genome DNA appears;
(7) centrifuging at 12,000rpm (13,400 Xg) for 5min, and discarding the supernatant;
(8) adding 200 μ l 70% ethanol, vortexing and shaking for 5s, centrifuging at 12,000rpm (13,400 Xg) for 2min, and discarding the supernatant;
(9) inverting the centrifuge tube on clean absorbent paper for at least 5min to ensure settling in the tube;
(10) air-drying the DNA precipitate until all liquid is completely volatilized;
(11) adding AE 50 μ l, storing in refrigerator at 4 deg.C for 10 days, detecting DNA concentration and purity with ultraviolet spectrometer, diluting according to the concentration, packaging, and storing in refrigerator at-20 deg.C.
3. The research respectively carries out whole exon sequencing on family members, is completed by Beijing Nuo He genesis science and technology Co., Ltd, and comprises the steps of detecting DNA samples; building a library for capturing; performing stock inspection; and (4) performing sequencing on the machine.
4. And (3) biological information analysis flow: as shown in fig. 3, including sequencing data quality assessment; detecting variation; screening variation and predicting disease correlation.
Screening the pathogenic genes according to conditions:
(1) sequencing the genome-wide exons of ALL people in the family, eliminating the mutations with MAF > 0.01 in ALL four databases of snp/Indel, ESP6500siv2-ALL, genomic AD-ALL and gnoma AD-EAS, and keeping 1882 snp/Indel mutation sites in which missense mutation, frameshift mutation and mutation generating stop codon are ALL contained;
(2) at least three predicted harmful Mutation sites in SIFT (< 0.05), Polyphen-2 (> 0.909), Mutation Taster (D) and CADD (> 20) software are reserved, and 1310 snp/Indel Mutation sites are reserved;
(3) according to the genetic model of autosomal dominant/sex, 161 snp/Indel variant sites are left after screening;
(4) the Phenolyzer software is used for predicting the correlation between the selected candidate genes and non-syndrome type congenital missing teeth, and the C.G103A mutation of the FGFR1 gene is an important mutation site which is likely to cause the non-syndrome type congenital missing teeth in the family through analysis.
To further verify the association of the c.g103a mutation in the third exon of FGFR1 gene with the non-syndromic congenital missing tooth, we further verified whether the family members carried the gene by Sanger sequencing, which was done by bio-engineering (shanghai) gmbh:
(1) extracting DNA and inspecting quality;
(2) primer designs (shown in SEQ ID NO: 1 and SEQ ID NO: 2) were designed for PCR using software Primer Premier 5;
(3) after PCR amplification, separating the PCR product by 1% agarose gel electrophoresis, and cutting and recovering the target PCR band;
(4) the sequencing product is purified, subjected to electrophoresis on a machine, and analyzed by sequence analysis, and the genotyping result shows that non-syndromic patients with congenital teeth deficiency are heterozygous mutations, while normal control shows no mutation at the site, as shown in figure 4. Therefore, according to the design scheme provided by the invention, the fact that the c.G103A mutation on the third exon of the FGFR1 gene detected by whole genome exon sequencing can be a new pathogenic site of non-syndrome type congenital missing tooth and can cause diseases by hybridization can be successfully confirmed in the embodiment, and therefore, the FGFR1 mutant gene provided by the invention can be used for non-syndrome type congenital missing tooth mechanism research, screening, auxiliary diagnosis and/or treatment.
Sequence listing
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gcggcgccgg ggcctccgca gggcgatgga gcccggtctg caaggaaagt gaggcgccgc 360
cgctgcgttc tggaggaggg gggcacaagg tctggagacc ccgggtggcg gacgggagcc 420
ctccccccgc cccgcctccg gggcaccagc tccggctcca ttgttcccgc ccgggctgga 480
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cgctcgcaca agccacggcg gactctcccg aggcggaacc tccacgccga gcgagggtca 660
gtttgaaaag gaggatcgag ctcactgtgg agtatccatg gagatgtgga gccttgtcac 720
caacctctaa ctgcagaact gggatgtgga gctggaagtg cctcctcttc tgggctgtgc 780
tggtcacagc cacactctgc accgctaggc cgtccccgac cttgcctgaa caagcccagc 840
cctggagagc ccctgtggaa gtggagtcct tcctggtcca ccccggtgac ctgctgcagc 900
ttcgctgtcg gctgcgggac gatgtgcaga gcatcaactg gctgcgggac ggggtgcagc 960
tggcggaaag caaccgcacc cgcatcacag gggaggaggt ggaggtgcag gactccgtgc 1020
ccgcagactc cggcctctat gcttgcgtaa ccagcagccc ctcgggcagt gacaccacct 1080
acttctccgt caatgtttca gatgctctcc cctcctcgga ggatgatgat gatgatgatg 1140
actcctcttc agaggagaaa gaaacagata acaccaaacc aaaccgtatg cccgtagctc 1200
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tcgaggtgaa tgggagcaag attggcccag acaacctgcc ttatgtccag atcttgaaga 1680
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ctagtttgct ttgctgacca aatgcctggt accagaggat ggtgaggcga aggccaggtt 3780
gggggcagtg ttgtggccct ggggcccagc cccaaactgg gggctctgta tatagctatg 3840
aagaaaacac aaagtgtata aatctgagta tatatttaca tgtcttttta aaagggtcgt 3900
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ctacttttgc tgttttattt ttttaaatta tgttctaaac ctattttcag tttaggtccc 4080
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cgcacctcct gcctctccgc tgcttttgga aagtcagaaa aagaagatgt ctgcttcgag 4320
ggcaggaacc ccatccatgc agtagaggcg ctgggcagag agtcaaggcc cagcagccat 4380
cgaccatgga tggtttcctc caaggaaacc ggtggggttg ggctggggag ggggcaccta 4440
cctaggaata gccacggggt agagctacag tgattaagag gaaagcaagg gcgcggttgc 4500
tcacgcctgt aatcccagca ctttgggaca ccgaggtggg cagatcactt caggtcagga 4560
gtttgagacc agcctggcca acttagtgaa accccatctc tactaaaaat gcaaaaatta 4620
tccaggcatg gtggcacacg cctgtaatcc cagctccaca ggaggctgag gcagaatccc 4680
ttgaagctgg gaggcggagg ttgcagtgag ccgagattgc gccattgcac tccagcctgg 4740
gcaacagaga aaacaaaaag gaaaacaaat gatgaaggtc tgcagaaact gaaacccaga 4800
catgtgtctg ccccctctat gtgggcatgg ttttgccagt gcttctaagt gcaggagaac 4860
atgtcacctg aggctagttt tgcattcagg tccctggctt cgtttcttgt tggtatgcct 4920
ccccagatcg tccttcctgt atccatgtga ccagactgta tttgttggga ctgtcgcaga 4980
tcttggcttc ttacagttct tcctgtccaa actccatcct gtccctcagg aacgggggga 5040
aaattctccg aatgtttttg gttttttggc tgcttggaat ttacttctgc cacctgctgg 5100
tcatcactgt cctcactaag tggattctgg ctcccccgta cctcatggct caaactacca 5160
ctcctcagtc gctatattaa agcttatatt ttgctggatt actgctaaat acaaaagaaa 5220
gttcaatatg ttttcatttc tgtagggaaa atgggattgc tgctttaaat ttctgagcta 5280
gggatttttt ggcagctgca gtgttggcga ctattgtaaa attctctttg tttctctctg 5340
taaatagcac ctgctaacat tacaatttgt atttatgttt aaagaaggca tcatttggtg 5400
aacagaacta ggaaatgaat ttttagctct taaaagcatt tgctttgaga ccgcacagga 5460
gtgtctttcc ttgtaaaaca gtgatgataa tttctgcctt ggccctacct tgaagcaatg 5520
ttgtgtgaag ggatgaagaa tctaaaagtc ttcataagtc cttgggagag gtgctagaaa 5580
aatataaggc actatcataa ttacagtgat gtccttgctg ttactactca aatcacccac 5640
aaatttcccc aaagactgcg ctagctgtca aataaaagac agtgaaattg a 5691
<210>4
<211>5691
<212>DNA
<213> human (Homo sapiens)
<400>4
gcatagcgct cggagcgctc ttgcggccac aggcgcggcg tcctcggcgg cgggcggcag 60
ctagcgggag ccgggacgcc ggtgcagccg caacgcgcgg aggaacccgg gtgtgccggg 120
agctgggcgg ccacgtccgg acgggaccga gacccctcgt agcgcattgc ggcgacctcg 180
ccttccccgg ccgcgagcgc gccgctgctt gaaaagccgc ggaacccaag gacttttctc 240
cggtccgagc tcggggcgcc ccgcagggcg cacggtaccc gtgctgcagt cgggcacgcc 300
gcggcgccgg ggcctccgca gggcgatgga gcccggtctg caaggaaagt gaggcgccgc 360
cgctgcgttc tggaggaggg gggcacaagg tctggagacc ccgggtggcg gacgggagcc 420
ctccccccgc cccgcctccg gggcaccagc tccggctcca ttgttcccgc ccgggctgga 480
ggcgccgagc accgagcgcc gccgggagtc gagcgccggc cgcggagctc ttgcgacccc 540
gccaggaccc gaacagagcc cgggggcggc gggccggagc cggggacgcg ggcacacgcc 600
cgctcgcaca agccacggcg gactctcccg aggcggaacc tccacgccga gcgagggtca660
gtttgaaaag gaggatcgag ctcactgtgg agtatccatg gagatgtgga gccttgtcac 720
caacctctaa ctgcagaact gggatgtgga gctggaagtg cctcctcttc tgggctgtgc 780
tggtcacagc cacactctgc accgctaggc cgtccccgac cttgcctgaa caagcccagc 840
cctggggagc ccctgtggaa gtggagtcct tcctggtcca ccccggtgac ctgctgcagc 900
ttcgctgtcg gctgcgggac gatgtgcaga gcatcaactg gctgcgggac ggggtgcagc 960
tggcggaaag caaccgcacc cgcatcacag gggaggaggt ggaggtgcag gactccgtgc 1020
ccgcagactc cggcctctat gcttgcgtaa ccagcagccc ctcgggcagt gacaccacct 1080
acttctccgt caatgtttca gatgctctcc cctcctcgga ggatgatgat gatgatgatg 1140
actcctcttc agaggagaaa gaaacagata acaccaaacc aaaccgtatg cccgtagctc 1200
catattggac atccccagaa aagatggaaa agaaattgca tgcagtgccg gctgccaaga 1260
cagtgaagtt caaatgccct tccagtggga ccccaaaccc cacactgcgc tggttgaaaa 1320
atggcaaaga attcaaacct gaccacagaa ttggaggcta caaggtccgt tatgccacct 1380
ggagcatcat aatggactct gtggtgccct ctgacaaggg caactacacc tgcattgtgg 1440
agaatgagta cggcagcatc aaccacacat accagctgga tgtcgtggag cggtcccctc 1500
accggcccat cctgcaagca gggttgcccg ccaacaaaac agtggccctg ggtagcaacg 1560
tggagttcat gtgtaaggtg tacagtgacc cgcagccgca catccagtgg ctaaagcaca 1620
tcgaggtgaa tgggagcaag attggcccag acaacctgcc ttatgtccag atcttgaaga 1680
ctgctggagt taataccacc gacaaagaga tggaggtgct tcacttaaga aatgtctcct 1740
ttgaggacgc aggggagtat acgtgcttgg cgggtaactc tatcggactc tcccatcact 1800
ctgcatggtt gaccgttctg gaagccctgg aagagaggcc ggcagtgatg acctcgcccc 1860
tgtacctgga gatcatcatc tattgcacag gggccttcct catctcctgc atggtggggt 1920
cggtcatcgt ctacaagatg aagagtggta ccaagaagag tgacttccac agccagatgg 1980
ctgtgcacaa gctggccaag agcatccctc tgcgcagaca ggtgtctgct gactccagtg 2040
catccatgaa ctctggggtt cttctggttc ggccatcacg gctctcctcc agtgggactc 2100
ccatgctagc aggggtctct gagtatgagc ttcccgaaga ccctcgctgg gagctgcctc 2160
gggacagact ggtcttaggc aaacccctgg gagagggctg ctttgggcag gtggtgttgg 2220
cagaggctat cgggctggac aaggacaaac ccaaccgtgt gaccaaagtg gctgtgaaga 2280
tgttgaagtc ggacgcaaca gagaaagact tgtcagacct gatctcagaa atggagatga 2340
tgaagatgat cgggaagcat aagaatatca tcaacctgct gggggcctgc acgcaggatg 2400
gtcccttgta tgtcatcgtg gagtatgcct ccaagggcaa cctgcgggag tacctgcagg 2460
cccggaggcc cccagggctg gaatactgct acaaccccag ccacaaccca gaggagcagc 2520
tctcctccaa ggacctggtg tcctgcgcct accaggtggc ccgaggcatg gagtatctgg 2580
cctccaagaa gtgcatacac cgagacctgg cagccaggaa tgtcctggtg acagaggaca 2640
atgtgatgaa gatagcagac tttggcctcg cacgggacat tcaccacatc gactactata 2700
aaaagacaac caacggccga ctgcctgtga agtggatggc acccgaggca ttatttgacc 2760
ggatctacac ccaccagagt gatgtgtggt ctttcggggt gctcctgtgg gagatcttca 2820
ctctgggcgg ctccccatac cccggtgtgc ctgtggagga acttttcaag ctgctgaagg 2880
agggtcaccg catggacaag cccagtaact gcaccaacga gctgtacatg atgatgcggg 2940
actgctggca tgcagtgccc tcacagagac ccaccttcaa gcagctggtg gaagacctgg 3000
accgcatcgt ggccttgacc tccaaccagg agtacctgga cctgtccatg cccctggacc 3060
agtactcccc cagctttccc gacacccgga gctctacgtg ctcctcaggg gaggattccg 3120
tcttctctca tgagccgctg cccgaggagc cctgcctgcc ccgacaccca gcccagcttg 3180
ccaatggcgg actcaaacgc cgctgactgc cacccacacg ccctccccag actccaccgt 3240
cagctgtaac cctcacccac agcccctgct gggcccacca cctgtccgtc cctgtcccct 3300
ttcctgctgg caggagccgg ctgcctacca ggggccttcc tgtgtggcct gccttcaccc 3360
cactcagctc acctctccct ccacctcctc tccacctgct ggtgagaggt gcaaagaggc 3420
agatctttgc tgccagccac ttcatcccct cccagatgtt ggaccaacac ccctccctgc 3480
caccaggcac tgcctggagg gcagggagtg ggagccaatg aacaggcatg caagtgagag 3540
cttcctgagc tttctcctgt cggtttggtc tgttttgcct tcacccataa gcccctcgca 3600
ctctggtggc aggtgccttg tcctcagggc tacagcagta gggaggtcag tgcttcgtgc 3660
ctcgattgaa ggtgacctct gccccagata ggtggtgcca gtggcttatt aattccgata 3720
ctagtttgct ttgctgacca aatgcctggt accagaggat ggtgaggcga aggccaggtt 3780
gggggcagtg ttgtggccct ggggcccagc cccaaactgg gggctctgta tatagctatg3840
aagaaaacac aaagtgtata aatctgagta tatatttaca tgtcttttta aaagggtcgt 3900
taccagagat ttacccatcg ggtaagatgc tcctggtggc tgggaggcat cagttgctat 3960
atattaaaaa caaaaaagaa aaaaaaggaa aatgttttta aaaaggtcat atattttttg 4020
ctacttttgc tgttttattt ttttaaatta tgttctaaac ctattttcag tttaggtccc 4080
tcaataaaaa ttgctgctgc ttcatttatc tatgggctgt atgaaaaggg tgggaatgtc 4140
cactggaaag aagggacacc cacgggccct ggggctaggt ctgtcccgag ggcaccgcat 4200
gctcccggcg caggttcctt gtaacctctt cttcctaggt cctgcaccca gacctcacga 4260
cgcacctcct gcctctccgc tgcttttgga aagtcagaaa aagaagatgt ctgcttcgag 4320
ggcaggaacc ccatccatgc agtagaggcg ctgggcagag agtcaaggcc cagcagccat 4380
cgaccatgga tggtttcctc caaggaaacc ggtggggttg ggctggggag ggggcaccta 4440
cctaggaata gccacggggt agagctacag tgattaagag gaaagcaagg gcgcggttgc 4500
tcacgcctgt aatcccagca ctttgggaca ccgaggtggg cagatcactt caggtcagga 4560
gtttgagacc agcctggcca acttagtgaa accccatctc tactaaaaat gcaaaaatta 4620
tccaggcatg gtggcacacg cctgtaatcc cagctccaca ggaggctgag gcagaatccc 4680
ttgaagctgg gaggcggagg ttgcagtgag ccgagattgc gccattgcac tccagcctgg 4740
gcaacagaga aaacaaaaag gaaaacaaat gatgaaggtc tgcagaaact gaaacccaga 4800
catgtgtctg ccccctctat gtgggcatgg ttttgccagt gcttctaagt gcaggagaac 4860
atgtcacctg aggctagttt tgcattcagg tccctggctt cgtttcttgt tggtatgcct 4920
ccccagatcg tccttcctgt atccatgtga ccagactgta tttgttggga ctgtcgcaga 4980
tcttggcttc ttacagttct tcctgtccaa actccatcct gtccctcagg aacgggggga 5040
aaattctccg aatgtttttg gttttttggc tgcttggaat ttacttctgc cacctgctgg 5100
tcatcactgt cctcactaag tggattctgg ctcccccgta cctcatggct caaactacca 5160
ctcctcagtc gctatattaa agcttatatt ttgctggatt actgctaaat acaaaagaaa 5220
gttcaatatg ttttcatttc tgtagggaaa atgggattgc tgctttaaat ttctgagcta 5280
gggatttttt ggcagctgca gtgttggcga ctattgtaaa attctctttg tttctctctg 5340
taaatagcac ctgctaacat tacaatttgt atttatgttt aaagaaggca tcatttggtg 5400
aacagaacta ggaaatgaat ttttagctct taaaagcatt tgctttgaga ccgcacagga 5460
gtgtctttcc ttgtaaaaca gtgatgataa tttctgcctt ggccctacct tgaagcaatg 5520
ttgtgtgaag ggatgaagaa tctaaaagtc ttcataagtc cttgggagag gtgctagaaa 5580
aatataaggc actatcataa ttacagtgat gtccttgctg ttactactca aatcacccac 5640
aaatttcccc aaagactgcg ctagctgtca aataaaagac agtgaaattg a 5691
Claims (8)
1. A non-syndromic congenital missing tooth related FGFR1 mutant gene is characterized in that the mutant FGFR1 is a heterozygous mutation, FGFR1: exon3: c.G103A; the transcriptome number of the wild FGFR1 gene in NCBI database is NM-001174063, the 103 th base of the mutant gene in the third exon of the wild transcript is mutated from G to A, and other parts are the same as the wild type.
2. A mutant FGFR1 protein, wherein the wild-type FGFR1 protein has the numbering in the NCBI database of: NP-001167534, wherein the amino acid at 35 th position of wild FGFR1 protein is mutated from glycine to arginine.
3. The FGFR1 mutant gene of claim 1, for use in research, screening, auxiliary diagnosis and/or treatment of non-syndromic congenital missing tooth mechanism.
4. A detection method for detecting FGFR1 mutant genes is characterized by comprising the following steps:
(1) extracting genome DNA;
(2) sanger sequencing: and (3) carrying out genotype detection on the FGFR1 mutant gene synthesis primer, wherein the 103 th base in the third exon of the FGFR1 gene sequence is mutated from G to A and is an indication of non-syndrome type congenital missing teeth.
5. The method of claim 4, wherein the primer sequence is as set forth in SEQ ID NO: 1 and SEQ ID NO: 2, respectively.
6. The method of claim 4, wherein the biological sample from which the genomic DNA is extracted is blood.
7. A primer for detecting the FGFR1 mutant gene has a sequence shown in SEQ ID NO: 1 and SEQ ID NO: 2, respectively.
8. Use of the primers of claim 7 in the preparation of a reagent for screening a FGFR1 mutant gene.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109897894A (en) * | 2018-12-27 | 2019-06-18 | 黄欢 | A kind of pathogenic mutation and its detection reagent of the infull disease of Osteogenic developmental |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109897894A (en) * | 2018-12-27 | 2019-06-18 | 黄欢 | A kind of pathogenic mutation and its detection reagent of the infull disease of Osteogenic developmental |
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| 丁婷婷等: "一个非综合征型先天缺牙家系的MSX1基因突变分析", 《山东大学学报(医学版)》 * |
| 杨艳等: "1例非综合征型先天缺牙家系的基因筛查", 《武汉大学学报(医学版)》 * |
| 谭灵等: "3例非综合征多数牙先天缺失家系的基因筛查", 《牙体牙髓牙周病学杂志》 * |
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