CN103160597A - Molecular biology method for predicating bouncing potentials of excellent ice-snow sportsmen - Google Patents
Molecular biology method for predicating bouncing potentials of excellent ice-snow sportsmen Download PDFInfo
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- CN103160597A CN103160597A CN2013101159326A CN201310115932A CN103160597A CN 103160597 A CN103160597 A CN 103160597A CN 2013101159326 A CN2013101159326 A CN 2013101159326A CN 201310115932 A CN201310115932 A CN 201310115932A CN 103160597 A CN103160597 A CN 103160597A
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Abstract
The invention discloses a molecular biology method for predicating the bouncing potentials of excellent ice-snow sportsmen, wherein the 577R alleles of ACTN3 (alpha-actinin-3 genes) genes are dominant alleles for strength quality and speed quality, various researches on a resistance training effect show that the training sensitivities of R allele carriers are higher, and the close relation between the polymorphisms of the ACTN3 genes and the strength quality is incontrovertible; additionally, I alleles in ACE (angiotensin-converting enzyme) genome may also be the genes related to the strength quality. According to the molecular biology method disclosed by the invention, the bouncing potentials of the ice-snow sportsmen are predicated by determining the R/X polymorphism of ACTN3 genotype and the I/D polymorphism of ACE genotype in the blood source genomic DNAs (deoxyribonucleic acid) of the ice-snow sportsmen, namely, ACTN3-RR type groups having the ACE genes which are homozygous insertion (II) type are selected. The molecular biology method disclosed by the invention can be used for early selection for the excellent ice-snow sportsmen.
Description
Technical field
The present invention is a kind of bounce molecular biology method of potential of outstanding winter sports person of predicting, namely uses molecular biology method that winter sports person's jump potential is predicted, belongs to the cross discipline of sports science and bio-science.
Background technology
Spring is winter sports person's an important special quality, and jump event requires the sportsmen to show at short notice top speed or peak power, and is high to the explosive power requirement, needs energy supply high-power, the short period of time.The training that traineres will bounce invariably is placed on the critical role of outstanding winter sports person's body building.Spring is a synthesis quality, and the development of spring is subjected to impact and the restriction of many factors.Although the motor capacity that the environmental factorss such as day after tomorrow training and nutrition also can the remarkably influenced human body is studied and is found that also athletic genomic constitution and polymorphism are determining their physical efficiency to a great extent.Explosive power has various definitions, is exactly briefly the power of human body, and namely strength multiply by speed, and a lot of winter sports projects all need the sportsmen that good spring is arranged.As figure skating, jump action has been most important composition in free skating, and various jump actions occupy great per-cent in free skating content now.As a sportsmen who takes part in game, the ability of completing single jump action is not singly arranged, and should be able to join jumping and jump continuously.Jump action is the result of muscle work done, be by the ice face upspring health, freely skim over aerial and drop to some displacements of sliding backward on ice.Their outstanding feature be the requirement sportsmen within the of short duration time, raise to greatest extent Muscle contraction strength, given play to the physical efficiency potentiality, the sportsmen need to have higher speed quality and Velocity-force.
Studies show that in a large number α-actin-3 (α-actinin-3, ACTN3) gene and human muscle's explosive power is closely related, the ACTN3 assignment of genes gene mapping has the conservative property of height in evolution on o.11 karyomit(e), all very important on the structure and function of mankind's muscle.Approximately 1,000,000,000 people lack ACTN3 albumen in the world.This is relevant with mankind ACTN3 gene polynorphisms.The 1747th of the 16th exon of ACTN3 gene wild-type (R) is C, just becomes mutant (X) after C has been mutated into T.Therefore when individual ACTN3 genotype was the 577XX homozygote, the 577th amino-acid residue (arginine) just become a stop code, thereby causes the disappearance of ACTN3.
Speed quality and neural speed of response, sensitivity, coordination and musculature, muscle fiber types are relevant, be subjected to the impact of congenital hereditary larger, develop critical porion early, therefore can use earlier the presentation index to carry out test evaluation, for early stage selection provides foundation.It is more late that Velocity-force develops critical porion, just can reach higher level to postpuberty.Therefore, utilize the ACTN3 gene level of the individual explosive power of Accurate Prediction in early days, can improve undoubtedly the success ratio of Velocity-force item group personnel training.Studies confirm that to have the advantage of explosive power with the allelic individuality of R, and with the allelic individuality of X no matter on explosive power or endurance, all do not show any special feature.The sports achievement that the sportsmen who carries this gene more easily obtains than the noncarrier when accepting the equal in quality training.New molecular biology evidence shows, the physiologically active owing to having improved the aerobic metabolism relevant enzymes has increased slow switch fibers quantity, has reduced the fast muscle fiber diameter, causes the muscle strength in ACTN3 knock-out mice body significantly to reduce.
It is relevant to human motion talent that angiotensin i-converting enzyme (angiotensin I converting enzyme, ACE) gene pleiomorphism also has been proved.The ACE gene is positioned at the 17q23 chromosomal region, total length 21kb, contain 26 exons and 25 introns, consist of insertion/deletion ((Insertion/Deletion, I/D) polymorphism of ACE gene at No. 16 intron as mark take the tumor-necrosis factor glycoproteins of one section 287bp.
All there were significant differences on genotype frequency or gene frequency (P＜0.02 and P＜0.003) no matter sportsmen and ordinary person are found in research, and the elite mostly is the ACE-II homozygote, and rare DD homozygote.Generally believe the I/D polymorphism that really there is the ACE gene and the dependency of motor capacity.One piece of result of study that Williams etc. are published on " Nature " magazine is thought, the ratio of muscle work done and energy expenditure (DE) is the best index of estimating muscle efficiency, its research find through after the training of 11 weeks only the genotypic DE of ACE-II significantly increase.Relative high economical energy expenditure state is revealed than ACE3/ID with DD genospecies body surface in another research report ACE-II genotype colony, and fat free body weight is also higher than other genotype.These results of study support that all I allelotrope is mainly to affect motor capacity by muscle efficiency.In addition, with the ACE level of outstanding endurance exercise person's colony's blood plasma of ACE-I allelic association and cardiac muscular tissue also higher than other genotype.
If a sportsmen has the assortment of genes of multiple promotion motor capacity simultaneously, he probably obtains than quantum jump in the field of oneself.Affect discovery and the Property Identification of the gene variant of motor capacity, make and carry out DNA tests in children, become possibility to pick out the talent who is fit to certain specific sports sports events or to optimize training method.
Summary of the invention
The present invention predicts winter sports person's spring potential by detecting ACTN3 gene R/X polymorphism and ACE gene I/D polymorphism.
Embodiment
Main agents:
Erythrocyte cracked liquid (10mmol/L Tris-HCL pH7.6,5mmol/L, MgCl
2, 10mmol/L NaCl), write cell lysis buffer (10mmol/L Tris, pH7.6,10mmol/L EDTA, 50mmol/L NaCl), Proteinase K, 10%SDS, 6M saturated sodium-chloride, phenol chloroform isoamyl alcohol (25: 24: 1).
Operation steps:
1, extract blood
Vein extracts winter sports person's venous blood 10mL, anticoagulant heparin.
2, the extraction of genomic dna and purifying
(1) with anticoagulation with the centrifugal 5min of 1500r/min, sucking-off blood plasma.
(2) add isopyknic erythrocyte cracked liquid, the abundant mixing that turns upside down, the centrifugal 15min of 2500～3000r/min removes supernatant liquor.
(3) add appropriate erythrocyte cracked liquid 2～3mL, the abundant mixing that turns upside down, the centrifugal 12min of 4000r/min removes supernatant liquor.
(4) repeating step is (3) 2～3 times, adds write cell lysis buffer 3mL, and the mixing that turns upside down adds appropriate 10%SDS, shakes up gently, adds 10mg/mL RNAase, hatches 2h in 37 ℃ of water-baths, adds the 10mg/mL Proteinase K, overnight incubation in 37 ℃ of waters bath with thermostatic control.
(5) add the saturated phenol of equivalent volumes, the centrifugal 10min of 2500r/min after mixing gets supernatant liquor (noting not being drawn onto the protein between organic phase and inorganic phase).
(6) add the phenol chloroform isoamyl alcohol (25: 24: 1) of equivalent volumes, the centrifugal 10min of 5000r/min gets the centrifugal 10min of chloroform isoamyl alcohol (24: 1) 5000r/min that supernatant liquor adds equivalent.
(7) get supernatant liquor, add the dehydrated alcohol of 2 times of volume precoolings, shake gently to cotton-shaped DNA and separate out, the centrifugal 5min of 1000～1500r/min makes the adherent ethanol that goes of DNA, cleans once with 70% ethanol, and the centrifugal 5min of 1000～1500r/min removes ethanol.
(8) appropriate 70% ethanol is resuspended with DNA, moves in the eppendof pipe of 0.15mL, and the centrifugal 5min of 1000～1500r/min removes ethanol, seasoning.
(9) DNA is dissolved in the TE damping fluid.Get 2 μ LDNA samples, with A (OD) value and the DNA concentration of ultraviolet spectrophotometry test sample, last DNA all is adjusted to 100 μ g/mL.
3, polymerase chain reaction (Polymerase Chain Reaction, PCR)
(1) design of primers
ACTN3 upstream region of gene primer: 5 ' CTG TTG CCT GTG GTA AGT GGG-3 '
ACTN3 gene downstream primer: 5 ' TGG TCA CAG TAT GCA GGA GGG-3 '
ACE upstream region of gene primer: 5 ' CTG GAG ACC ACT CCC ATC CTT TCT-3 '
ACE gene downstream primer: 5 ' GAT GTG GCC ATC ACA TTC GTC AGA T-3 '
(2) ACTN3 gene-PCR reaction system and loop parameter
PCR reaction system: 3 μ L ddH
2O, upstream and downstream primer 1 μ L, 10 μ L 2 * Reddy Mix (5mM MgCl
2), 5 μ L genomic dnas.
The gene PCR reaction conditions: 95 ℃ of denaturation 5min, 94 ℃ of sex change 30s, 60 ℃ of annealing 30s, 72 ℃ are extended 30s, totally 35 circulations, 72 ℃ are extended 30s, and the extension circulation is complete, takes out the PCR pipe, is placed in 4 ℃ of refrigerators to be measured.
(3) ACE gene-PCR reaction system and loop parameter
PCR reaction system: total system 25 μ L, ddH
2O15 μ L, dNTP2.0 μ L, 10*buffer2.5 μ L (include Mg
2+, concentration is 15nm/L), Mg
2+1.0 μ L (concentration 25nm/L), primer each 1.0 μ L, template 2.0 μ L Taq enzyme 0.5 μ L.
ACE gene PCR reaction conditions: 94 ℃ of denaturation 4min, 94 ℃ of sex change 1min, 55.5 ℃ of annealing 1min, 72 ℃ are extended 1min30s, totally 30 circulations, 72 ℃ are extended 10min, and the extension circulation is complete, takes out the PCR pipe, is placed in 4 ℃ of refrigerators to be measured.
4, ACTN3 and ACE genotype identification
(1) ACTN3 genotype identification
Get 15 μ L PCR products and mix with Loadingbuffer, 3% agarose gel electrophoresis is 90min approximately, takes out gel piece after electrophoresis and is positioned in ultraviolet analyzer, observes electrophoretic band and uses the camera picture.The genotypic individuality of RR according to MARKER identification 86,97,108 and 205bp size fragment.
(2) ACE genotype identification
Agarose gel electrophoresis: by 2: 7 volumes, Loadingbuffer is mixed with the PCR product, 1% agarose gel electrophoresis is 40min approximately, takes out gel piece after electrophoresis and is positioned in ultraviolet analyzer, observes electrophoretic band and uses the camera picture.Screening only has the homozygote insert type (II) of 401bp band.
5, statistics screening
Judge the genotype of person's to be measured ACTN3 gene and ACE gene according to the stripe size of gel electrophoresis.Filtering out the ACTN3 gene is that RR type while ACE gene is the individuality of II type.
Claims (2)
1. the present invention is a kind of bounce molecular biology method of potential of outstanding winter sports person of predicting.
2. according to method claimed in claim 1, it is characterized in that predicting by measuring ACTN3 genotype R/X polymorphism and ACE genotype I/D polymorphism in the genomic dna of winter sports person's blood source winter sports person's spring potential.Namely screen ACTN3 (RR) type while ACE gene and be the individuality of (II) type.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105861731A (en) * | 2016-06-14 | 2016-08-17 | 杭州势必健生物科技有限公司 | Detection kit for genes related to sport advantages of Chinese people and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1732270A (en) * | 2002-09-16 | 2006-02-08 | 遗传技术有限公司 | ACTN3 genotype screen for athletic performance |
| CN102864223A (en) * | 2012-08-30 | 2013-01-09 | 山东百福基因科技有限公司 | Detection kit for highly specific multiple genes related to movement functions and detection method thereof |
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2013
- 2013-04-07 CN CN2013101159326A patent/CN103160597A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1732270A (en) * | 2002-09-16 | 2006-02-08 | 遗传技术有限公司 | ACTN3 genotype screen for athletic performance |
| CN102864223A (en) * | 2012-08-30 | 2013-01-09 | 山东百福基因科技有限公司 | Detection kit for highly specific multiple genes related to movement functions and detection method thereof |
Non-Patent Citations (2)
| Title |
|---|
| 杨贤罡等: "《ACTN3基因R577X多态性与运动能力的关联性研究：Meta分析》", 《体育科学》 * |
| 许汪宇等: "《ACTN3基因多态性与肌肉爆发力及基因选材的关系：Meta分析》", 《中国优生优育》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105861731A (en) * | 2016-06-14 | 2016-08-17 | 杭州势必健生物科技有限公司 | Detection kit for genes related to sport advantages of Chinese people and application thereof |
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Application publication date: 20130619 |