CN106399500A - Molecular identification method of Protosalanx hyalocranius and neosalanx taihuensis - Google Patents
Molecular identification method of Protosalanx hyalocranius and neosalanx taihuensis Download PDFInfo
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- 241000969498 Neosalanx taihuensis Species 0.000 title claims abstract description 39
- 238000000034 method Methods 0.000 title claims abstract description 14
- 241001039489 Protosalanx hyalocranius Species 0.000 title abstract 8
- 230000002068 genetic effect Effects 0.000 claims abstract description 18
- 238000012408 PCR amplification Methods 0.000 claims abstract description 8
- 101150001086 COB gene Proteins 0.000 claims abstract description 6
- 101150053771 MT-CYB gene Proteins 0.000 claims abstract description 6
- 101150006264 ctb-1 gene Proteins 0.000 claims abstract description 6
- 101150088166 mt:Cyt-b gene Proteins 0.000 claims abstract description 6
- 238000012163 sequencing technique Methods 0.000 claims abstract description 6
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims description 10
- 241000969495 Hemisalanx Species 0.000 claims description 9
- 108010075028 Cytochromes b Proteins 0.000 claims description 8
- 239000003153 chemical reaction reagent Substances 0.000 claims description 7
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 6
- 108010010803 Gelatin Proteins 0.000 claims description 5
- 239000008273 gelatin Substances 0.000 claims description 5
- 229920000159 gelatin Polymers 0.000 claims description 5
- 235000019322 gelatine Nutrition 0.000 claims description 5
- 235000011852 gelatine desserts Nutrition 0.000 claims description 5
- 229910001629 magnesium chloride Inorganic materials 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 108090000790 Enzymes Proteins 0.000 claims description 3
- 102000004190 Enzymes Human genes 0.000 claims description 3
- 238000000137 annealing Methods 0.000 claims description 3
- 238000006243 chemical reaction Methods 0.000 claims description 3
- 230000007850 degeneration Effects 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims description 3
- 238000000746 purification Methods 0.000 claims description 3
- 238000011144 upstream manufacturing Methods 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 abstract description 3
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- PHTQWCKDNZKARW-UHFFFAOYSA-N isoamylol Chemical compound CC(C)CCO PHTQWCKDNZKARW-UHFFFAOYSA-N 0.000 description 4
- 241000251468 Actinopterygii Species 0.000 description 2
- 102100025287 Cytochrome b Human genes 0.000 description 2
- 241001062511 Salangidae Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 235000019688 fish Nutrition 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000012850 discrimination method Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 229960004756 ethanol Drugs 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- LCCNCVORNKJIRZ-UHFFFAOYSA-N parathion Chemical compound CCOP(=S)(OCC)OC1=CC=C([N+]([O-])=O)C=C1 LCCNCVORNKJIRZ-UHFFFAOYSA-N 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000003252 repetitive effect Effects 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
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Abstract
The invention discloses a molecular identification method of Protosalanx hyalocranius and neosalanx taihuensis. The method for molecular genetic identification of Protosalanx hyalocranius and neosalanx taihuensis comprises (1) respectively extracting the genome DNAs of Protosalanx hyalocranius and neosalanx taihuensis to be identified, (2) carrying out PCR amplification on Cytb genes of Protosalanx hyalocranius and neosalanx taihuensis through the extracted genome DNAs as templates, and (3) purifying the amplified product and carrying out direct sequencing, wherein the Protosalanx hyalocranius has a basic group T at the 45th site of the Ctyb gene and the neosalanx taihuensis has a basic group C at the 45th site of the Ctyb gene. The method can quickly, easily, accurately and effectively identify Protosalanx hyalocranius and neosalanx taihuensis. The results are stable and a repetition rate is high. The method fills the gaps of the current domestic molecular biological standard identification of Protosalanx hyalocranius and neosalanx taihuensis.
Description
Technical field
The invention belongs to genetic arts, it is related to the molecular identification method of a kind of Big Salangid and neosalanx taihuensis.
Background technology
China is the area of origin of world Hemisalanx and main areal area, extensively divides in each big water system in Eastern China and lake
Cloth.Hemisalanx nutritive value and economic worth are all very high, are important economic fishs, in creating huge economic benefit.Hemisalanx
Biocycle is short, discrete of generation, reproductivity and to settle down ability strong.As typical r- Kstrategist, Hemisalanx is sensitive to environmental change
And be swift in response, Population Dynamic is fast.But the Hemisalanx natural resources of China but because reclaiming land from a lake, overfishing, environmental pollution and
The impact of the many factors such as Habitat Fragmentation and continuous downturn, the natural resources of various Hemisalanxes all decline to some extent, species
Distribution is reduced significantly, and indivedual species are gradually endangered, and carries out Icefish Resource investigation and protection work is extremely urgent.Big Salangid and Taihu Lake
New Hemisalanx is two kinds of common Hemisalanx species, both body colours and plesiomorphism, is difficult to distinguish and differentiates.
Content of the invention
The purpose of the present invention is the difficulty for differentiating forms, provides the molecular genetic of a kind of Big Salangid and neosalanx taihuensis
Discrimination method.
The purpose of the present invention is achieved through the following technical solutions：
A kind of Big Salangid and the molecular genetic identification method of neosalanx taihuensis, comprise the steps of：
(1) Big Salangid to be identified and neosalanx taihuensis genomic DNA are extracted respectively；
(2) with the genomic DNA of extraction as template, PCR expands the cytochrome b gene of Big Salangid and neosalanx taihuensis
(Ctyb)；
(3) amplified production after purification, direct Sequencing, Cytb gene the 45th site base be T for Big Salangid, if this site
It is C for neosalanx taihuensis.
The forward primer of the described cytochrome b gene expanding Big Salangid and neosalanx taihuensis for PCR is L14321：
SEQ ID NO.1, downstream primer is H15634：SEQ ID NO.2.
Described PCR reaction system is 50 μ L：The template DNA 1 μ L of 100ng/ μ L, PCR buffer 5 μ L, dNTP mixed liquor
4 μ L, each 1 μ L of upstream and downstream primer of every kind of dNTP 0.1mmol/L, 10 μm of ol/L, the Taq enzyme of 2 μ L 2.5IU；Distilled water 36 μ
L；Described PCR buffer is by 10mmol/L Tris-HCl, pH9.0,0.5mmol/L KCl, 30mmol/L MgCl2,
0.01% (g/100ml) gelatin forms；Pcr amplification reaction program is：94 DEG C of denaturations 4min, 94 DEG C of degeneration 40s, 55 DEG C of annealing
40s, 72 DEG C extension 90s, through 35 circulation after again 72 DEG C extension 10min.
The primer pair that a kind of molecular genetic for Big Salangid and neosalanx taihuensis differentiates, forward primer is L14321：SEQ
ID NO.1, downstream primer is H15634：SEQ ID NO.2.
Primer pair of the present invention differentiates the application in Big Salangid and neosalanx taihuensis in molecular genetic.
Application in preparing Big Salangid and neosalanx taihuensis molecular genetic identification reagent for the primer pair of the present invention.
A kind of Big Salangid and neosalanx taihuensis molecular genetic identification reagent, comprise primer pair of the present invention.
Described Big Salangid and neosalanx taihuensis molecular genetic identification reagent, further preferably include PCR buffer and dNTP mixes
Close liquid, described PCR buffer is by 10mmol/L Tris-HCl, pH9.0,0.5mmol/L KCl, 30mmol/L MgCl2,
0.01% gelatin composition.
Beneficial effect：
The present invention is directed to Big Salangid and neosalanx taihuensis cytochrome b (Cytochrome b, Cytb) gene order difference,
First using the cytochrome b gene full length sequence of Big Salangid and neosalanx taihuensis as foundation, thus qualitative identification Big Salangid and
Neosalanx taihuensis.The method can quickly, easy, accurately and efficiently differentiate Big Salangid and neosalanx taihuensis, result stability
Good, repetitive rate is high, has filled up the blank that current domestic standard molecular biological differentiates Big Salangid and neosalanx taihuensis, has been also beneficial to
Icefish Resource is protected.
Brief description
Fig. 1 is Cytb gene sequencing sequence alignment result in the present invention, and what asterisk was indicated is from the 45th site (Cytb-45)
The base sequence difference starting.
Specific embodiment
To be further elucidated with the present invention with reference to embodiments, but to be not that the present invention is limited in any form, only
Only illustrate.
Embodiment 1
1st, design of primers
Design a pair of specific PCR amplimer, primer sequence is：L14321：5'-CCAGTGA-
CTTGAAAAACCACCG-3'(SEQ ID NO.1), downstream primer is H15634：5'-CTTAGCTTTGGGAGT-TAAGGGT-
3'(SEQ ID NO.2).
2nd, sample collection
Take Big Salangid to be identified and individual each 30 tails of neosalanx taihuensis.
3rd, extracting genome DNA
The tail fin taking every tail fish organizes about 30mg, blots surface moisture with filter paper, loads 1.5ml centrifuge tube, adds 420 μ L
STE buffer (30mmol/L Tris-HCl, pH8.0,200mmol/L EDTA, 50mmol/L NaCl).Add after mixing
Concentration is the E.C. 3.4.21.64 10 μ L of 10% SDS 80 μ L and 20mg/mL, 56 DEG C of digestion 8-10h；Add equal-volume phenol：Chloroform：
Isoamyl alcohol (volume ratio 25:24:1) twice, 12000r is centrifuged 10min, takes supernatant for extracting；Add equal-volume chloroform：Isoamyl alcohol
(volume ratio 24:1) once, 12000r is centrifuged 10min, takes supernatant for extracting；Add equal-volume pre-cooling dehydrated alcohol, 12000r from
Heart 10min, abandons clear liquid；Add the washing with alcohol precipitation of 600mL 70%, 8000r is centrifuged 10min, outwells ethanol, is dried；Add
200 μ L distilled water dissolvings, ultraviolet spectrophotometer is surveyed its OD value and is diluted to 100ng/ μ L, and -20 DEG C save backup.
4th, PCR amplification and detection
The DNA of extraction is template, enters performing PCR amplification with above-mentioned primer (L14321 and H15634).PCR reaction system is
50μL：1 μ L template DNA (100ng/ μ L)；PCR buffer 5 μ L (10mmol/L Tris-HCl, pH9.0,0.5mmol/LKCl,
30mmol/L MgCl2, 0.01% gelatin)；DNTP mixed liquor 4 μ L (every kind of dNTP 0.1mmol/L)；Upstream and downstream primer (10 μ
Mol/L) each 1 μ L, 2 μ L Taq enzyme (2.5IU)；Distilled water 36 μ L.Amplified reaction program is：94 DEG C of denaturations 4min, 94 DEG C of changes
Property 40s, 55 DEG C annealing 40s, 72 DEG C extension 90s, through 35 circulation after again 72 DEG C extension 10min.
PCR primer detects in 1% agarose gel electrophoresiies and separates, and after electrophoresis 30min under 120V voltage, becomes through gel
As analysis system determines size and the purity of pcr amplification product.
5th, sequencing compares
Pcr amplification product after detection is fed directly to biotech firm's purification sequencing, obtains 1141bp sequence, permissible after comparison
Find that the base sequence from the 45th site (Cytb-45) for the Cytb gene has differences：30 tail Big Salangid the 45th site (Cytb-45)
Base is T, and 30 tail neosalanx taihuensis the 45th site (Cytb-45) is C.
Embodiment 2
According to embodiment 1 method, by sample enlargement to Big Salangid and individual each 80 tails of neosalanx taihuensis, result still shows
Big Salangid cytochrome b gene the 45th site (Cytb-45) base is T, and neosalanx taihuensis the 45th site (Cytb-45) base is
C.
Claims (8)
1. a kind of Big Salangid and the molecular genetic identification method of neosalanx taihuensis are it is characterised in that comprise the steps of：
(1) Big Salangid to be identified and neosalanx taihuensis genomic DNA are extracted respectively；
(2) with the genomic DNA of extraction as template, PCR expands the cytochrome b gene Ctyb of Big Salangid and neosalanx taihuensis；
(3) amplified production after purification, direct Sequencing, Cytb gene the 45th site base be T for Big Salangid, if this site is C
For neosalanx taihuensis.
2. molecular genetic identification method according to claim 1 it is characterised in that for PCR amplification Big Salangid new with Taihu Lake
The forward primer of the cytochrome b gene of Hemisalanx is L14321：SEQ ID NO.1, downstream primer is H15634：SEQ ID
NO.2.
3. molecular genetic identification method according to claim 1 is it is characterised in that PCR reaction system is 50 μ L：100ng/μ
The template DNA 1 μ L of L, PCR buffer 5 μ L, dNTP mixed liquor 4 μ L, every kind of dNTP 0.1mmol/L, 10 μm of ol/L's is upper and lower
The trip each 1 μ L of primer, the Taq enzyme of 2 μ L 2.5IU；Distilled water 36 μ L；Described PCR buffer by 10mmol/L Tris-HCl,
PH9.0,0.5mmol/L KCl, 30mmol/L MgCl2, 0.01% (w/v) gelatin composition.
4. molecular genetic identification method according to claim 1 is it is characterised in that pcr amplification reaction program is：94 DEG C pre-
Degeneration 4min, 94 DEG C of degeneration 40s, 55 DEG C annealing 40s, 72 DEG C extension 90s, through 35 circulation after again 72 DEG C extension 10min.
5. a kind of primer pair differentiates the application in Big Salangid and neosalanx taihuensis in molecular genetic, and described primer pair is drawn by upstream
Thing is L14321：SEQ ID NO.1, downstream primer is H15634：SEQ ID NO.2 forms.
6. application in preparing Big Salangid and neosalanx taihuensis molecular genetic identification reagent for a kind of primer pair, described primer pair
It is L14321 by forward primer：SEQ ID NO.1, downstream primer is H15634：SEQ ID NO.2 forms.
7. a kind of Big Salangid and neosalanx taihuensis molecular genetic identification reagent are it is characterised in that comprise forward primer L14321：SEQ
ID NO.1 and downstream primer H15634：The primer pair of SEQ ID NO.2 composition.
8. Big Salangid according to claim 7 and neosalanx taihuensis molecular genetic identification reagent are it is characterised in that described
Big Salangid and neosalanx taihuensis molecular genetic identification reagent, further preferably include PCR buffer and dNTP mixed liquor, described PCR
Buffer is by 10mmol/L Tris-HCl, pH9.0,0.5mmol/L KCl, 30mmol/L MgCl2, 0.01% gelatin composition.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107858437A (en) * | 2017-09-27 | 2018-03-30 | 浙江海洋大学 | A kind of molecular biology method of quick discriminating silver color schizothoracin and Yi Li schizothoracins |
| CN110699460A (en) * | 2019-09-23 | 2020-01-17 | 江苏省淡水水产研究所 | Cytb gene sequence based molecular identification method for identifying beta and beta |
| CN113913534A (en) * | 2021-11-19 | 2022-01-11 | 中国水产科学研究院黑龙江水产研究所 | Primer for identifying big whitebait and Taihu new whitebait and identification method thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104152451A (en) * | 2014-08-19 | 2014-11-19 | 淮南师范学院 | Primer and method for molecular identification of new whitebait species in Taihu Lake |
-
2016
- 2016-09-18 CN CN201610829411.0A patent/CN106399500B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104152451A (en) * | 2014-08-19 | 2014-11-19 | 淮南师范学院 | Primer and method for molecular identification of new whitebait species in Taihu Lake |
Non-Patent Citations (2)
| Title |
|---|
| JIE ZHANG等: "molecular phylogeny of icefish salangidae based on complete mtDNA cytochrome b sequences, with comments on estuarine fish evolution", 《BIOLOGICAL JOURNAL OF THE LINNEAN SOCIETY》 * |
| 李大命等: "太湖大银鱼（protosalanx chinensis）细胞色素b基因序列多态性分析", 《江苏农业学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107858437A (en) * | 2017-09-27 | 2018-03-30 | 浙江海洋大学 | A kind of molecular biology method of quick discriminating silver color schizothoracin and Yi Li schizothoracins |
| CN110699460A (en) * | 2019-09-23 | 2020-01-17 | 江苏省淡水水产研究所 | Cytb gene sequence based molecular identification method for identifying beta and beta |
| CN113913534A (en) * | 2021-11-19 | 2022-01-11 | 中国水产科学研究院黑龙江水产研究所 | Primer for identifying big whitebait and Taihu new whitebait and identification method thereof |
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