CN106480046A - GNAS gene mutation body and its application - Google Patents

GNAS gene mutation body and its application Download PDF

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CN106480046A
CN106480046A CN201610925155.5A CN201610925155A CN106480046A CN 106480046 A CN106480046 A CN 106480046A CN 201610925155 A CN201610925155 A CN 201610925155A CN 106480046 A CN106480046 A CN 106480046A
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gnas gene
gnas
primer
gene
dna
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唐雨晨
郑芬萍
林细华
潘倩倩
李霖
李红
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Zhejiang University ZJU
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    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

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Abstract

The invention belongs to field of pharmaceutical biology, it is related to GNAS gene mutation body, a kind of for the family examination of autosomal dominant disorder and the GNAS gene mutation body of prenatal gene examination and its application.The present invention provides a kind of nucleic acid of detached GNAS gene mutation body, GNAS gene (the SEQ ID NO:1) c.310delG lack.In addition present invention also offers a kind of method of detection GNAS gene mutation body.Advantages of the present invention and effect：The method is easy, it is only necessary to pointedly detect, to patient family member, the mutation having been found that, you can diagnose pseudohypoparathyroidism from molecular biology angle.Fetus to patient carries out Prenatal Screening, can instruct aristogenesis.

Description

GNAS gene mutation body and its application
Technical field
The invention belongs to field of pharmaceutical biology, is related to GNAS gene mutation body, a kind of family for autosomal dominant disorder The GNAS gene mutation body of examination and prenatal gene examination and its application.
Background technology
GNAS gene is located at the long-armed of No. 20 chromosomes of the mankind, is a compound imprinted gene, can transcribe generation multiple Gene outcome.Most important of which product is the stimulatory G protein alpha subunit (Gs α) of its 1-13 exon coding.G-protein It is a kind of immanent signal conductive protein, multiple hormones can be mediated by generation second messenger's CAMP (cAMP) and send out The effect of waving.The GNAS gene mutation of coding Gs α can cause Gs alpha expression to reduce and/or function reduction causes Albright heredity Property osteodystrophy (AHO).While GNAS or tissue-specific imprinted gene, in proximal tubular, thyroid gland, hypophysis With mainly express maternal allele in ovary tissue.The mutation of maternity causes AHO phenotype and multiple hormones by first shape Parathyrine (PTH), thyrotropic hormone (TSH) and promoting sexual gland hormone opposing, i.e. Albright's syndrome to type Ia type.And it is paternal The mutation in source only results in AHO phenotype, i.e. vacation-Albright's syndrome to type[1].
GNAS gene mutation is distributed in whole Gs α coding and montage region, and nearly 50% is the disappearance for causing frameshift mutation Or insertion mutation, other mutation also include missense mutation, nonsense mutation, splice site mutation, in-frame deletion or insertion and complete Portion or partial gene deletion.The 4bp disappearance for being wherein located at the codon 189-190 position of exon 7 is a hot spot mutation.In Asia Research in crowd is simultaneously few, mutation type is based on frameshift mutation and missense mutation, it has been found that a small amount of nonsense mutation, montage Site mutation and full gene disappearance[1].
PCR is polymerase chain reaction, and the technology is in template DNA, in the presence of primer and four kinds of deoxyribonucleotides, according to The enzyme' s catalysis in archaeal dna polymerase are relied to react.Archaeal dna polymerase with single stranded DNA as template, by artificial synthesized antisense oligonucleotide primer Thing is combined with one section of complementary series in single-stranded DNA templates, forming part double-strand.At suitable temperature and environment, DNA is polymerized Deoxymononucleotide is added to 3 '-OH end of primer by enzyme, and as starting point, is extended along 5 ' → 3 ' direction of template, synthesis one The new DNA complementary strand of bar.Touchdown PCR (touchdown PCR) is a kind of round pcr, is mainly used in the optimization of the condition of PCR. The design of primer causes PCR to be difficult in many cases, such as specificity not enough easily mispairing etc..Touchdown PCR provides one Individual more easy optimization method, its principle is substantially：Expand at a higher temperature first, although now amplification efficiency is low, But non-specific amplification does not have substantially.With the reduction of annealing temperature, non-specific amplification progressively can increase.But due to now special Amplified production has reached certain predominance, therefore can produce strong Competitive assays to non-specific amplification, so as to significantly Improve the specificity and efficiency of PCR.From the beginning of higher than 5 DEG C of Tm value, each circulation successively decreases 1 DEG C to annealing temperature, until being less than Tm value 5 DEG C, general with 10 circulations, afterwards with less than 5 DEG C of Tm value as annealing temperature, then carry out 20-25 and circulate.
Gene sequencing Introduction on Principle：DNA double Sanger dideoxy DNA sequencing is that Sanger was initiated equal to 1977, and the technology is using double de- Oxygen ribonucleoside triphosphote (ddNTP) is used as sequencing reaction chain terminating agent.DdNTP is with common deoxyribonucleoside triphosphate (dNTP) no It is that they lack a hydroxyl at 3 ' ends of deoxyribose with part.They can be held by 5 ' under archaeal dna polymerase effect Triphosphoric acid group is mixed in the normal DNA for extending.But due to not having 3 ' hydroxyls, they can not form phosphoric acid with follow-up dNTP Diester linkage, therefore, the DNA for extending can not continue to extend and terminate.The examination used of 3100 sequenator of American AB I PRISM Include 4 kinds of fluorescent markers in agent BigDye, 4 kinds of marks are connected with 4 kinds of ddNTP respectively.Therefore, formed in sequencing reaction with The nucleic acid fragment mixture that machine terminates.The mixture through the Capillary Electrophoresis on sequenator, by the laser scanning of special wavelength After computer disposal, each base signal on nucleotide chain is showed with collection of illustrative plates form, here it is seen by us Sequencing chromatogram.
Content of the invention
This research needs to find trouble by genes of interest DNA extraction, design of primers, touchdown PCR and gene sequencing technology GNAS gene mutation site and type in disease individuality, so as to instruct clinician to carry out same gene position to patient family member The examination of point, and instruct prenatal gene examination.
The invention provides a kind of nucleic acid of detached GNAS gene mutation body and its polypeptide.
A kind of nucleic acid of detached GNAS gene mutation body, GNAS gene (the SEQ ID NO:1) c.310delG lack (i.e. the 53rd G base deletion of the 4th extron, sequence are SEQ ID NO:2).
A kind of detached polypeptide, the polypeptide are GNAS gene (SEQ ID NO:1) nucleic acid coding for c.310delG lacking (sequence be SEQ ID NO:4).
In addition present invention also offers a kind of method of detection GNAS gene mutation body.
A kind of method that non-diagnostic purpose detects the GNAS gene mutation body, including expanded using following primer Step：(sequence is SEQ ID NO to upstream primer 5 '-TACTCCTAACTGACATGGTG-3 ':5)；Downstream primer 5 '- (sequence is SEQ ID NO to ATATGGACACTGTGCTCAGG-3 ':6).
Specifically include following steps：
(1) extraction of genomic DNA
Whole blood is extracted using poba gene group DNA extraction system (0.1-20ml) RelaxGene Blood DNA System Genomic DNA；
(2) the PCR amplification of GNAS gene 1-13 exon
1) PCR reaction system
GNAS gene 1-13 exon primer sequence
25 μ l systems:
2) PCR response procedures (1-13 exon)
(3) detection of PCR primer, recovery and sequencing
PCR primer is detected using 1% agarose gel electrophoresis, is returned in strict accordance with Tiangeng plain agar sugar gel DNA The operating procedure for receiving kit carries out PCR primer purifying, and send Hangzhou Qing Ke Zi Xi Bioisystech Co., Ltd to be sequenced.
Further, present invention also offers the biology of the screening pseudohypoparathyroidism of non-diagnostic purpose The method of sample, comprises the steps：
Sample of nucleic acid is extracted from biological sample；
The nucleotide sequence of the sample of nucleic acid is determined by said method, and is detected；
If it is the biological sample or biological sample that c.310delG the GNAS gene of the nucleotide sequence of the nucleic acid samples lacks Product source and its family member are susceptible to suffer from the instruction of pseudohypoparathyroidism.
In addition, the invention provides detection kit.
A kind of kit for screening the biological sample of pseudohypoparathyroidism, comprising detection GNAS gene C.310delG, the reagent of mutant, the GNAS gene are lacked.
A kind of kit for fetus Prenatal Screening, the reagent comprising detection GNAS gene mutation body, the GNAS base Because c.310delG lacking.
Wherein, the reagent is nucleic acid probe.
Described kit (or nucleic acid probe) includes one group of primer pair, 5 '-TACTCCTAACTGACATGGTG- of upstream primer 3’；Downstream primer 5 '-ATATGGACACTGTGCTCAGG-3 '.
In addition the present invention provides kit answering in the examination of pseudohypoparathyroidism family and Prenatal Screening With.
The present invention is by whole blood sample collection, design of primers, the extraction of genomic DNA, GNAS gene 1-13 exon PCR amplification, the detection of PCR primer, recovery and sequencing obtain GNAS gene mutation result：
GNAS gene 1-13 exon (including sub-piece comprising be close to) all amplifies target stripe (accompanying drawing 1).Logical Cross Chromas and DNAMAN software sequencing result is compared, find patient's GNAS the 53rd G base of the 4th extron Disappearance causes frameshift mutation, according to the method for expressing that gives tacit consent in the world, is expressed as c.310delG (accompanying drawing 2).
Prediction (DNAMAN software and Swiss are simulated by amino acid and protein structure that mutant nucleotide sequence is encoded Model online homologous modeling (http://swissmodel.expasy.org//SWISS-MODEL.html)[3], find patient The deletion mutation c.310delG of GNAS gene may cause protein translation frameshit, and may cause the of GNAS gene code 104 glutamic acid are changed into lysine, and occur at the 110th amino acids to terminate (accompanying drawing 3) in advance, it is possible to create truncated protein Or it is degraded (accompanying drawing 4).
Advantages of the present invention and effect：The method is easy, it is only necessary to patient family member is pointedly detected and is had been found that Mutation, you can from molecular biology angle diagnose pseudohypoparathyroidism.Fetus to patient carries out antenatal sieve Look into, aristogenesis can be instructed.
Description of the drawings
Fig. 1 is that GNAS gene 1-13 exon expands electrophoretogram.Band is from left to right followed successively by exons 1, and 2,3,4/ 5,6,7/8,9/10,11/12,13, negative control, molecular marked compound.
Fig. 2 patient (a) and 4/5 gene order of GNAS extron of normal person (b).A in figure occurs in that two kinds of sequences, one Kind is normal sequence, and another kind of is mutant nucleotide sequence.As arrow pointed location G (black signal) is lacked, the A in next site is (green Chrominance signal) occur in advance, i.e., each nucleotides after mutational site is advanced by a site.
The normal GNAS of Fig. 3 and the corresponding amino acid sequence of mutation GNAS coded sequence.
After normal Gs α protein three-dimensional structure (left figure) and mutation after the homologous modeling of Fig. 4, Gs α protein three-dimensional structure is (right Figure).
Specific embodiment
The present invention is expanded on further with reference to specific embodiment, it should be appreciated that following examples are merely to illustrate the present invention Rather than limit the scope of the invention.
The material wanted needed for the embodiment of the present invention, reagent except specified otherwise all can market buy.The method for being used is non- Specified otherwise is this area conventional method.
The invention provides a kind of nucleic acid of detached GNAS gene mutation body and its polypeptide.
The nucleic acid of detached GNAS gene mutation body, GNAS gene (the SEQ ID NO:1) c.310delG lack (i.e. The 4th G base deletion of extron the 53rd, sequence are SEQ ID NO:2).
GNAS gene 1-13 exon (including sub-piece comprising be close to) all amplifies target stripe (accompanying drawing 1).Logical Cross Chromas and DNAMAN software sequencing result is compared, find patient's GNAS the 53rd G base of the 4th extron Disappearance causes frameshift mutation, according to the method for expressing that gives tacit consent in the world, is expressed as c.310delG (accompanying drawing 2).
Normal GNAS gene nucleotide series：(NCBI reference sequences NM_000516.4)
Mutation GNAS gene nucleotide series：
A kind of detached polypeptide, the polypeptide are GNAS gene (SEQ ID NO:1) nucleic acid coding for c.310delG lacking (sequence be SEQ ID NO:4).
Prediction (DNAMAN software and Swiss are simulated by amino acid and protein structure that mutant nucleotide sequence is encoded Model online homologous modeling (http://swissmodel.expasy.org//SWISS-MODEL.html)[3], find patient The deletion mutation c.310delG of GNAS gene may cause protein translation frameshit, and may cause the of GNAS gene code 104 glutamic acid are changed into lysine, and occur at the 110th amino acids to terminate (accompanying drawing 3) in advance, it is possible to create truncated protein Or it is degraded (accompanying drawing 4).And then judge whether to be susceptible to suffer from pseudohypoparathyroidism.
The amino acid sequence of normal GNAS gene code：
The amino acid sequence of mutation GNAS gene code：
The invention provides the method for detection GNAS gene mutation body, comprises the steps：
(1) whole blood sample collection
Whole blood (being provided by the first affiliated hospital of Zhejiang University) loads containing EDTANa4Anticoagulant tube, be put into one 80 DEG C of refrigerators Preserve, in case applying when extracting DNA.
(2) design of primers
Primer is with reference to existing document.According to GNAS gene order (Genebank ID：NT 011362), application Primmer5 Software separately designs specific primer to GNAS gene 1-13 exon (including sub-piece comprising be close to), application Each region that methylates of MethPrimer software separately designs two pairs of primers, Methylation-specific primer MF/MR and non-methylates Specific primer UF/UR, all primers are synthesized by Hangzhou Qing Ke Zi Xi Bioisystech Co., Ltd.Primer is referring to table 1.1[2].
1.1 GNAS gene 1-13 exon primer sequence of table
(3) extraction of genomic DNA
Whole blood is extracted using poba gene group DNA extraction system (0.1-20ml) RelaxGene Blood DNA System Genomic DNA, is carried out to specifications.
(4) the PCR amplification of GNAS gene 1-13 exon
1) PCR reaction system
25 μ l systems:
2) PCR response procedures (1-13 exon)
(5) detection of PCR primer, recovery and sequencing
PCR primer is detected using 1% agarose gel electrophoresis, is returned in strict accordance with Tiangeng plain agar sugar gel DNA The operating procedure for receiving kit carries out PCR primer purifying, and send Hangzhou Qing Ke Zi Xi Bioisystech Co., Ltd to be sequenced.
The present invention is also provided according to detection GNAS gene mutation body, and then screens the life of pseudohypoparathyroidism The method of thing sample.
(1) whole blood sample collection
The whole blood (being provided by the first affiliated hospital of Zhejiang University) of pseudohypoparathyroidism patient and normal person Load containing EDTANa4Anticoagulant tube, be put into one 80 DEG C of Refrigerator stores, in case extract DNA when apply.
(2) design of primers
Primer is with reference to existing document.According to GNAS gene order (Genebank ID：NT 011362), application Primmer5 Software separately designs specific primer to GNAS gene 1-13 exon (including sub-piece comprising be close to), application Each region that methylates of MethPrimer software separately designs two pairs of primers, Methylation-specific primer MF/MR and non-methylates Specific primer UF/UR, all primers are synthesized by Hangzhou Qing Ke Zi Xi Bioisystech Co., Ltd.Primer is referring to table 1.1[2].
1.1 GNAS gene 1-13 exon primer sequence of table
(3) extraction of genomic DNA
Whole blood is extracted using poba gene group DNA extraction system (0.1-20ml) RelaxGene Blood DNA System Genomic DNA, is carried out to specifications.
(4) the PCR amplification of GNAS gene 1-13 exon
1) PCR reaction system
25 μ l systems:
2) PCR response procedures (1-13 exon)
(5) detection of PCR primer, recovery and sequencing
PCR primer is detected using 1% agarose gel electrophoresis, is returned in strict accordance with Tiangeng plain agar sugar gel DNA The operating procedure for receiving kit carries out PCR primer purifying, and send Hangzhou Qing Ke Zi Xi Bioisystech Co., Ltd to be sequenced.
GNAS gene 1-13 exon (including sub-piece comprising be close to) all amplifies target stripe (accompanying drawing 1).Logical Cross Chromas and DNAMAN software sequencing result is compared, find patient's GNAS the 53rd G base of the 4th extron Disappearance causes frameshift mutation, according to the method for expressing that gives tacit consent in the world, is expressed as c.310delG (accompanying drawing 2).
Bibliography
[1]Turan S,Bastepe M.The GNAS complex locus and human diseases associated with loss-of-function mutations or epimutations within this imprinted gene.Hormone research in paediatrics 2013；80:229-41.
[2]Zhang XL.Analysis of Pseudohypoparathyroidism and GNAS1Gene[D] .Zhengzhou:Zhengzhou University,2011:21
[3]Arnold K.,Bordoli L.,Kopp J.,and Schwede T.(2006).The SWISS-MODEL Workspace:A web-based environment for protein structure homology modeling.Bioinformatics,22,195-201.
SEQUENCE LISTING
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<213>Mankind Homo sapiens (human)
<400> 3
Met Gly Cys Leu Gly Asn Ser Lys Thr Glu Asp Gln Arg Asn Glu Glu
1 5 10 15
Lys Ala Gln Arg Glu Ala Asn Lys Lys Ile Glu Lys Gln Leu Gln Lys
20 25 30
Asp Lys Gln Val Tyr Arg Ala Thr His Arg Leu Leu Leu Leu Gly Ala
35 40 45
Gly Glu Ser Gly Lys Ser Thr Ile Val Lys Gln Met Arg Ile Leu His
50 55 60
Val Asn Gly Phe Asn Gly Glu Gly Gly Glu Glu Asp Pro Gln Ala Ala
65 70 75 80
Arg Ser Asn Ser Asp Gly Glu Lys Ala Thr Lys Val Gln Asp Ile Lys
85 90 95
Asn Asn Leu Lys Glu Ala Ile Glu Thr Ile Val Ala Ala Met Ser Asn
100 105 110
Leu Val Pro Pro Val Glu Leu Ala Asn Pro Glu Asn Gln Phe Arg Val
115 120 125
Asp Tyr Ile Leu Ser Val Met Asn Val Pro Asp Phe Asp Phe Pro Pro
130 135 140
Glu Phe Tyr Glu His Ala Lys Ala Leu Trp Glu Asp Glu Gly Val Arg
145 150 155 160
Ala Cys Tyr Glu Arg Ser Asn Glu Tyr Gln Leu Ile Asp Cys Ala Gln
165 170 175
Tyr Phe Leu Asp Lys Ile Asp Val Ile Lys Gln Ala Asp Tyr Val Pro
180 185 190
Ser Asp Gln Asp Leu Leu Arg Cys Arg Val Leu Thr Ser Gly Ile Phe
195 200 205
Glu Thr Lys Phe Gln Val Asp Lys Val Asn Phe His Met Phe Asp Val
210 215 220
Gly Gly Gln Arg Asp Glu Arg Arg Lys Trp Ile Gln Cys Phe Asn Asp
225 230 235 240
Val Thr Ala Ile Ile Phe Val Val Ala Ser Ser Ser Tyr Asn Met Val
245 250 255
Ile Arg Glu Asp Asn Gln Thr Asn Arg Leu Gln Glu Ala Leu Asn Leu
260 265 270
Phe Lys Ser Ile Trp Asn Asn Arg Trp Leu Arg Thr Ile Ser Val Ile
275 280 285
Leu Phe Leu Asn Lys Gln Asp Leu Leu Ala Glu Lys Val Leu Ala Gly
290 295 300
Lys Ser Lys Ile Glu Asp Tyr Phe Pro Glu Phe Ala Arg Tyr Thr Thr
305 310 315 320
Pro Glu Asp Ala Thr Pro Glu Pro Gly Glu Asp Pro Arg Val Thr Arg
325 330 335
Ala Lys Tyr Phe Ile Arg Asp Glu Phe Leu Arg Ile Ser Thr Ala Ser
340 345 350
Gly Asp Gly Arg His Tyr Cys Tyr Pro His Phe Thr Cys Ala Val Asp
355 360 365
Thr Glu Asn Ile Arg Arg Val Phe Asn Asp Cys Arg Asp Ile Ile Gln
370 375 380
Arg Met His Leu Arg Gln Tyr Glu Leu Leu
385 390
<210> 4
<211> 109
<212> PRT
<213>Mankind Homo sapiens (human)
<400> 4
Met Gly Cys Leu Gly Asn Ser Lys Thr Glu Asp Gln Arg Asn Glu Glu
1 5 10 15
Lys Ala Gln Arg Glu Ala Asn Lys Lys Ile Glu Lys Gln Leu Gln Lys
20 25 30
Asp Lys Gln Val Tyr Arg Ala Thr His Arg Leu Leu Leu Leu Gly Ala
35 40 45
Gly Glu Ser Gly Lys Ser Thr Ile Val Lys Gln Met Arg Ile Leu His
50 55 60
Val Asn Gly Phe Asn Gly Glu Gly Gly Glu Glu Asp Pro Gln Ala Ala
65 70 75 80
Arg Ser Asn Ser Asp Gly Glu Lys Ala Thr Lys Val Gln Asp Ile Lys
85 90 95
Asn Asn Leu Lys Glu Ala Ile Lys Pro Leu Trp Pro Pro
100 105
<210> 5
<211> 20
<212> DNA
<213>Mankind Homo sapiens (human)
<400> 5
tactcctaac tgacatggtg 20
<210> 6
<211> 20
<212> DNA
<213>Mankind Homo sapiens (human)
<400> 6
atatggacac tgtgctcagg 20

Claims (10)

1. a kind of nucleic acid of detached GNAS gene mutation body, it is characterised in that：C.310delG, the GNAS gene is lacked.
2. a kind of detached polypeptide, it is characterised in that：The polypeptide is had the right nucleic acid coding described in requirement 1.
3. a kind of method that test right requires GNAS gene mutation body described in 1, it is characterised in that：Including being entered using following primer The step of row amplification：Upstream primer 5 '-TACTCCTAACTGACATGGTG-3 '；Downstream primer 5 '- ATATGGACACTGTGCTCAGG-3’.
4. method according to claim 3, it is characterised in that：Specifically include following steps：
(1) extraction of genomic DNA
Whole blood gene is extracted using poba gene group DNA extraction system (0.1-20ml) RelaxGene Blood DNA System Group DNA；
(2) the PCR amplification of GNAS gene 1-13 exon
1) PCR reaction system
25 μ l systems:
DEPC water 8μl Upstream primer (10 μM) 1.25μl Downstream primer (10 μM) 1.25μl 2×Taq PCR Master Mix 12.5μl Template DNA 2μl
2) PCR response procedures (1-13 exon)
(3) detection of PCR primer, recovery and sequencing
PCR primer is detected using 1% agarose gel electrophoresis, reclaims in strict accordance with Tiangeng plain agar sugar gel DNA and try The operating procedure of agent box carries out PCR primer purifying, and is sequenced, and judges that c.310delG GNAS gene lacks.
5. a kind of screening pseudohypoparathyroidism biological sample method, it is characterised in that：Comprise the steps：
Sample of nucleic acid is extracted from biological sample；
The nucleotide sequence of the sample of nucleic acid is determined by the method described in claim 3-4, and is detected；
If it is that the biological sample or biological sample come that c.310delG the GNAS gene of the nucleotide sequence of the nucleic acid samples lacks Source and its family member are susceptible to suffer from the instruction of pseudohypoparathyroidism.
6. a kind of kit for screening the biological sample of pseudohypoparathyroidism, it is characterised in that：Comprising inspection The reagent of GNAS gene mutation body is surveyed, c.310delG the GNAS gene lacks.
7. a kind of kit for fetus Prenatal Screening, it is characterised in that：Reagent comprising detection GNAS gene mutation body, institute State GNAS gene c.310delG to lack.
8. the kit according to claim 5 or 6, it is characterised in that：The reagent is nucleic acid probe.
9. the kit according to claim 5 or 6, it is characterised in that：Including one group of primer pair, upstream primer 5 '- TACTCCTAACTGACATGGTG-3’；Downstream primer 5 '-ATATGGACACTGTGCTCAGG-3 '.
10. kit described in claim 6-9 in the examination of pseudohypoparathyroidism family and Prenatal Screening should With.
CN201610925155.5A 2016-10-30 2016-10-30 GNAS gene mutation body and its application Pending CN106480046A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107904293A (en) * 2017-12-26 2018-04-13 南昌艾迪康医学检验实验室有限公司 Detect the primer and method of GNAS gene point mutations

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107904293A (en) * 2017-12-26 2018-04-13 南昌艾迪康医学检验实验室有限公司 Detect the primer and method of GNAS gene point mutations

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Application publication date: 20170308