CN102776284B - The diagnostic primers of Dai-ju stock Pseudosciaena crocea and Fujian-East Guangdong race large yellow croaker and discrimination method thereof - Google Patents
The diagnostic primers of Dai-ju stock Pseudosciaena crocea and Fujian-East Guangdong race large yellow croaker and discrimination method thereof Download PDFInfo
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Abstract
The invention discloses diagnostic primers and the discrimination method thereof of a kind of Dai-ju stock Pseudosciaena crocea and Fujian-East Guangdong race large yellow croaker, feature is primer sequence is primer pair J1:5-GGCATTTAGGCACAAGGAC-3,5-GGGGGCACAACTCAGAGG-3; Primer pair J2?: 5-CAGGCCCCACCATACTAGT-3,5-GGGTGCCGGTTCCTGAC-3, discrimination method is specific as follows: the step of large yellow croaker total DNA extraction and detection by quantitative; The step of preparation PCR amplification system and amplified reaction; Finally get product and carry out agarose gel electrophoresis detection, if 300-400bp band that is interval and 600-700bp interval all occurs in collection of illustrative plates, then this DNA sample is from Dai-ju stock Pseudosciaena crocea; Otherwise from Fujian-East Guangdong race large yellow croaker, it is high, reproducible that advantage has stability, differentiate that speed is fast, false positive rate is low.
Description
Technical field
The present invention relates to biotechnology species and differentiate field, especially relate to diagnostic primers and the discrimination method thereof of a kind of Dai-ju stock Pseudosciaena crocea and Fujian-East Guangdong race large yellow croaker.
Background technology
Large yellow croaker (
pseudosciaenacrocea), have another name called yellow croaker, yellow croaker, being under the jurisdiction of Perciformes (Perciformes), Sciaenidae (Sciaenidae), yellow croaker belongs to (Pseudosciaena), is one of China's marine fishing Main Commercial Fishes, is the maximum fish species of Chinese seawater cage cultured output.China coast large yellow croaker can be roughly divided into tai-chu race, Fujian-East Guangdong race and island in Guangdong Province race 3 geographical population.Because of the advantage in Dai-ju stock Pseudosciaena crocea is in body colour, build and local flavor etc., commercially price is higher than Fujian-East Guangdong race large yellow croaker; Dai-ju stock Pseudosciaena crocea body colour shows slightly golden yellow, and Fujian-East Guangdong race large yellow croaker body colour is lighter; On build, Dai-ju stock Pseudosciaena crocea build is taller and thinner, and Fujian-East Guangdong race large yellow croaker is then slightly fat; In nutritive value and local flavor, in Dai-ju stock Pseudosciaena crocea muscle tissue, the Major Nutrient value index of the reflection such as total amino acid content, essential amino acid, Fresh ear field and lipid acid meat flavor is all significantly higher than Fujian-East Guangdong race official well ocean family large yellow croaker.In addition, Dai-ju stock Pseudosciaena crocea occupies clear superiority than Fujian-East Guangdong race large yellow croaker in the speed of growth, surviving rate, sickness rate and feed coefficient etc.In Ningbo, the large yellow croaker overwhelming majority of Zhoushan Region cultivation is Fujian-East Guangdong race large yellow croaker, and it is very similar to Dai-ju stock Pseudosciaena crocea in appearance, adopts traditional character observation, and subjectivity by force, is also difficult to effectively distinguish.Therefore, exploitation Dai-ju stock Pseudosciaena crocea is different from the discrimination method of Fujian-East Guangdong race large yellow croaker, all seems extremely important for aspects such as plasm resource protection, fine-variety breeding and market sales.
Existing Chinese patent name is called that the diagnostic primers of different population large yellow croaker and discrimination method (application number is CN200610135259.2) disclose tai-chu race, the diagnostic primers of east Guangdong and island in Guangdong Province race large yellow croaker and discrimination method, adopt single primer RAPD-PCR AFLP system just can distinguish Dai-ju stock Pseudosciaena crocea and Fujian-East Guangdong race large yellow croaker, but, RAPD-PCR technology has very high requirement to the quality of DNA profiling, the condition of amplification and system, and RAPD technology self exists system instability, repeated low shortcoming.
Summary of the invention
Technical problem to be solved by this invention is to provide diagnostic primers and the discrimination method thereof of high, the reproducible Dai-ju stock Pseudosciaena crocea of a kind of stability and Fujian-East Guangdong race large yellow croaker.
The present invention solves the problems of the technologies described above adopted technical scheme: the diagnostic primers of a kind of Dai-ju stock Pseudosciaena crocea and Fujian-East Guangdong race large yellow croaker, and primer sequence is:
J1 upstream primer 5 '-GGCATTTAGGCACAAGGAC-3 ',
J1 downstream primer 5 '-GGGGGCACAACTCAGAGG-3 ';
J2 upstream primer 5 '-CAGGCCCCACCATACTAGT-3 ',
J2 downstream primer 5 '-GGGTGCCGGTTCCTGAC-3 '.
A discrimination method for Dai-ju stock Pseudosciaena crocea and Fujian-East Guangdong race large yellow croaker, concrete steps are as follows:
(1) large yellow croaker total DNA extraction and detection by quantitative
(2) PCR amplification system is prepared
Being formulated as follows of pcr amplification reaction system: various composition and final concentration are respectively: damping fluid 10mM, dNTP0.2mM, Mg
2+2mM, each primer is 0.2mM, Taq enzyme 1U, large yellow croaker template DNA 10-50ng, and with distilled water polishing to 25ul, described primer sequence is
J1 upstream primer 5 '-GGCATTTAGGCACAAGGAC-3 ',
J1 downstream primer 5 '-GGGGGCACAACTCAGAGG-3 ';
J2 upstream primer 5 '-CAGGCCCCACCATACTAGT-3 ',
J2 downstream primer 5 '-GGGTGCCGGTTCCTGAC-3 ';
(3) pcr amplification reaction
In 95 DEG C of heating 2-5min in PCR instrument, make the abundant sex change of template DNA, then enter amplification cycles, 94 DEG C of 30sec, 53-58 DEG C of 40sec, 72 DEG C of 35sec, carry out 27-30 circulation, finally, keep 5min, 4-16 DEG C of preservation at 72 DEG C;
(4) agarose gel electrophoresis detects
After pcr amplification reaction terminates, get product and carry out agarose gel electrophoresis detection, and observe with ultraviolet gel imaging system, take a picture, contrast according to electrophoresis result and the standard diagram provided, if 1 band all appears in 300-400bp interval and 600-700bp interval in collection of illustrative plates, then judge that this DNA sample is from Dai-ju stock Pseudosciaena crocea; If there is no band or only have a band, then judge that this DNA sample is from Fujian-East Guangdong race large yellow croaker.
In the pcr amplification reaction system of primer pair J1 large yellow croaker template DNA 10ng, primer pair J2 pcr amplification reaction system in large yellow croaker DNA40ng, PCR response procedures be: 95 DEG C of 3min, amplification cycles 94 DEG C of 30sec, 55.8 DEG C of 40sec, 72 DEG C of 35sec totally 28 circulations, 72 DEG C of 5min, 4 DEG C of preservations.Amplified production electrophorogram band can be made brighter.
PCR reaction terminate after, get product 10ul with 1.5% agarose gel electrophoresis detection.
Phenol chloroform extraction method is adopted to extract large yellow croaker DNA.
Compared with prior art, the invention has the advantages that: the diagnostic primers of Dai-ju stock Pseudosciaena crocea of the present invention and Fujian-East Guangdong race large yellow croaker and discrimination method thereof, diagnostic primers is used to carry out PCR reaction to J1 and J2, after electrophoresis is taken pictures, if sample somatotype is D1-N2 type (Fig. 2), D2-1 type (Fig. 3) or D2-2 type (Fig. 4), then determine that it is Fujian-East Guangdong race large yellow croaker; If sample OTU somatotype is D1-N1(and Fig. 1), then judge that this sample is as Dai-ju stock Pseudosciaena crocea, it is high, reproducible that above-mentioned discrimination method has stability, differentiates the advantages such as speed is fast, false positive rate is low.
Accompanying drawing explanation
Fig. 1 is the PCR primer electrophoretogram of D1-N1 type large yellow croaker;
Fig. 2 is the PCR primer electrophoretogram of D1-N2 type large yellow croaker;
Fig. 3 is the PCR primer electrophoretogram of D2-1 type large yellow croaker;
Fig. 4 is the PCR primer electrophoretogram of D2-2 type large yellow croaker.
Embodiment
Below in conjunction with accompanying drawing embodiment, the present invention is described in further detail.
Specific embodiment one
The diagnostic primers of a kind of Dai-ju stock Pseudosciaena crocea of the present invention and Fujian-East Guangdong race large yellow croaker, primer sequence is as follows:
J1 upstream primer 5 '-GGCATTTAGGCACAAGGAC-3 ',
J1 downstream primer 5 '-GGGGGCACAACTCAGAGG-3 ';
J2 upstream primer 5 '-CAGGCCCCACCATACTAGT-3 ',
J2-downstream primer 5 '-GGGTGCCGGTTCCTGAC-3 '.
The principle of invention: with large yellow croaker quick evolving region 2 pairs of amplimers; B1 upstream primer 5 '-AGACCTTCTAGGCTTTGCAATC-3 ', B1 downstream primer 5 '-GGAGCTTTCTAGGGCTCATCT-3 '; B2 upstream primer 5 '-TTATTTTCTATTCTACACCCTGGC-3 ', B2 downstream primer 5 '-AGAGGGAATACCCCGCTGT-3 ' carry out pcr amplification to Dai-ju stock Pseudosciaena crocea and Fujian-East Guangdong race large yellow croaker quick evolving region, the laggard row data analysis of cloning and sequencing.Adopt the B1 amplification mitochondrial D-loop region of large yellow croaker, to 2 partial sequence fragment binding analysis in this sequence, detect 2 OTU altogether, be respectively D1 type: GGCATTTAGGCACAAGGTC and CTTCTGAGTTGTGCCCCC, D2 type: GGCATTTAGGCACAAGGTT and TTTCTGAGTTGTGCCCCC; On the basis of above-mentioned experiment, use the primer pair B2 amplification mitochondrial ND4 region of large yellow croaker, to the 2 partial sequence fragments also binding analysis in this sequence, detect 2 OTU, be respectively N1 type: CAGGCCCCACCATACTAAT and GACAGGAACCGGCACCC, N2 type: CAGGCCCCACCATACTAAC and AACAGGAACCGGCACCC; Binding analysis is carried out to the OTU somatotype in above-mentioned D-loop and ND4 region, 3 types can be divided into: D1-N1, D1-N2 and D2.Dai-ju stock Pseudosciaena crocea is D1-N1 type; And 2 types can be divided in Fujian-East Guangdong race large yellow croaker: D1-N2 type and D2 type, D2 type has 2 kinds of collection of illustrative plates to be subdivided into D2-1 and D2-2 type again.
The efficiency of several base pair pcr amplifications of primer 3 ' end is most important.Mispairing 1 base, pcr amplification efficiency there will be certain decline, and mispairing 2 bases, and pcr amplification efficiency there will be and significantly declines.The present invention is when designing Dai-ju stock Pseudosciaena crocea and Fujian-East Guangdong race large yellow croaker diagnostic primers, and at above-mentioned sequence each fragment 3 ' end, the 2nd bit base place reciprocal introduces a mispairing.Make primer J1 upstream primer and J1 downstream primer like this, have 1 mispairing with D1 type DNA sample in 3 ' end second-to-last base respectively, with D2 type DNA sample in 3 ' least significant end mispairing, 2 bases; Also make primer J2 upstream primer and J2 downstream primer, have 1 mispairing with N1 type DNA sample in 3 ' end second-to-last base respectively, with N2 type DNA sample in 3 ' least significant end mispairing, 2 bases.
As shown in the figure, use diagnostic primers J1 and J2 to carry out PCR reaction, after electrophoresis is taken pictures, if sample somatotype is D1-N2 type (Fig. 2), D2-1 type (Fig. 3) or D2-2 type (Fig. 4), then DNA sample is from Fujian-East Guangdong race large yellow croaker; If sample OTU somatotype is D1-N1(and Fig. 1), then this DNA sample is from Dai-ju stock Pseudosciaena crocea.
Specific embodiment two
The discrimination method of a kind of Dai-ju stock Pseudosciaena crocea of the present invention and Fujian-East Guangdong race large yellow croaker, concrete steps are as follows:
(1) large yellow croaker total DNA extraction and detection by quantitative
(2) PCR amplification system is prepared
Being formulated as follows of pcr amplification reaction system: various composition and final concentration are respectively: damping fluid 10mM, dNTP0.2mM, Mg
2+2mM, the equal 0.2mM of each primer, Taq enzyme 1U, large yellow croaker template DNA 10-50ng, with distilled water polishing to 25ul, described primer sequence is:
J1 upstream primer 5 '-GGCATTTAGGCACAAGGAC-3 '
J1 downstream primer 5 '-GGGGGCACAACTCAGAGG-3 '
J2 upstream primer 5 '-CAGGCCCCACCATACTAGT-3 '
J2 downstream primer 5 '-GGGTGCCGGTTCCTGAC-3 '
(3) pcr amplification reaction
In 95 DEG C of heating 2-5min in PCR instrument, make the abundant sex change of template DNA, then enter amplification cycles, 94 DEG C of 30sec, 53-58 DEG C of 40sec, 72 DEG C of 35sec, carry out 27-30 circulation, finally, keep 5min, 4-16 DEG C of preservation at 72 DEG C;
(4) agarose gel electrophoresis detects
After pcr amplification reaction terminates, get product and carry out agarose gel electrophoresis detection, and observe with ultraviolet gel imaging system, take a picture, contrast according to electrophoresis result and the standard diagram provided, if 300-400bp is interval in collection of illustrative plates, the band in 600-700bp interval all occurs, then this DNA sample is from Dai-ju stock Pseudosciaena crocea; If do not have band or only have a band, then this DNA sample is from Fujian-East Guangdong race large yellow croaker.
Specific embodiment three
The discrimination method of a kind of Dai-ju stock Pseudosciaena crocea of the present invention and Fujian-East Guangdong race large yellow croaker, concrete steps are as follows:
(1) phenol chloroform extraction method is adopted to extract large yellow croaker DNA and detection by quantitative
(2) PCR amplification system is prepared
Being formulated as follows of pcr amplification reaction system: various composition and final concentration are respectively: damping fluid 10mM, dNTP0.2mM, Mg
2+2mM, in the pcr amplification reaction system of the equal 0.2mM of each primer, Taq enzyme 1U, primer pair J1 large yellow croaker template DNA 10ng, primer pair J2 pcr amplification reaction system in large yellow croaker DNA40ng, with distilled water polishing to 25ul, described primer sequence is
J1 upstream primer 5 '-GGCATTTAGGCACAAGGAC-3 '
J1 downstream primer 5 '-GGGGGCACAACTCAGAGG-3 '
J2 upstream primer 5 '-CAGGCCCCACCATACTAGT-3 '
J2 downstream primer 5 '-GGGTGCCGGTTCCTGAC-3 '
(3) pcr amplification reaction
PCR response procedures is: 95 DEG C of 3min, amplification cycles 94 DEG C of 30sec, 55.8 DEG C of 40sec, 72 DEG C of 35sec totally 28 circulations, 72 DEG C of 5min, 4 DEG C of preservations;
(4) agarose gel electrophoresis detects
After pcr amplification reaction terminates, get product 10ul, agarose gel electrophoresis with 1.5% detects, and observe with ultraviolet gel imaging system, take a picture, contrast according to electrophoresis result and the standard diagram provided, in collection of illustrative plates, 300-400bp is interval, and the band in 600-700bp interval all occurs, then this DNA sample is from Dai-ju stock Pseudosciaena crocea.
Consider practicality and the operability of the discrimination method of Dai-ju stock Pseudosciaena crocea.Find out a set of easy normalizing operation step and condition most important.Two PCR of the most key is preferred J1 and J2 react the response procedures and annealing temperature that carry out simultaneously.Real-timePCR technology has energy real-time quantitative to go out the advantage of the product of pcr amplification, and therefore this research adopts technique to be optimized terms and conditions.Same reaction system is adopted to carry out Real-timePCR discovery, when annealing temperature 55.8 DEG C, J1 is than the Ct value large 2 of J2, therefore the template DNA amount in the former PCR reaction system of strengthening is to 40ng, the pcr amplification of two pairs of primers can roughly synchronously be carried out, and pcr amplification cycle number is set to 28, the band that now D1, N1 type large yellow croaker sample P CR is increased is brighter, and the product of D2, N2 type pcr amplification is also failed detected by gel imaging system.
Use Real-timePCR to be optimized amplification condition, make pcr amplification effectively can distinguish D1 and D2 type, N1 and N2 type.For the Ct value of different annealing temperature, primer pair and template DNA sample in table 1, annealing temperature preferably 55.8 DEG C.Same reaction system is adopted to carry out Real-timePCR discovery, when annealing temperature 55.8 DEG C, J1 is than the Ct value large 2 of J2, and the template DNA amount in the former PCR reaction system of therefore strengthening is to 40ng, the pcr amplification of two pairs of primers can roughly synchronously be carried out, cycle number then preferably 28.Therefore the condition of PCR is found out, the band of D1 type and N1 type can be increased to can take pictures out with gel imaging system, D2 type and N2 type then can't detect band, then differentiate Dai-ju stock Pseudosciaena crocea and Fujian-East Guangdong race large yellow croaker according to the picture of gel imaging system.
The Ct value of table 1 different annealing temperature, primer pair and template DNA sample
Certainly, above-mentioned explanation is not limitation of the present invention, and the present invention is also not limited to above-mentioned citing.Those skilled in the art are in essential scope of the present invention, and the change made, remodeling, interpolation or replacement, also should belong to protection scope of the present invention.
Claims (5)
1. a diagnostic primers for Dai-ju stock Pseudosciaena crocea and Fujian-East Guangdong race large yellow croaker, is characterized in that primer sequence is:
J1 upstream primer 5 '-GGCATTTAGGCACAAGGAC-3 ',
J1 downstream primer 5 '-GGGGGCACAACTCAGAGG-3 ';
J2 upstream primer 5 '-CAGGCCCCACCATACTAGT-3 ',
J2 downstream primer 5 '-GGGTGCCGGTTCCTGAC-3 '.
2. a discrimination method for Dai-ju stock Pseudosciaena crocea and Fujian-East Guangdong race large yellow croaker, is characterized in that step is as follows:
(1) large yellow croaker total DNA extraction and detection by quantitative
(2) PCR amplification system is prepared
Being formulated as follows of pcr amplification reaction system: various composition and final concentration are respectively: damping fluid 10mM, dNTP0.2mM, Mg
2+2mM, each primer is 0.2mM, Taq enzyme 1U, large yellow croaker template DNA 10-50ng, and with distilled water polishing to 25ul, described primer sequence is
J1 upstream primer 5 '-GGCATTTAGGCACAAGGAC-3 ',
J1 downstream primer 5 '-GGGGGCACAACTCAGAGG-3 ';
J2 upstream primer 5 '-CAGGCCCCACCATACTAGT-3 ',
J2 downstream primer 5 '-GGGTGCCGGTTCCTGAC-3 '.
(3) pcr amplification reaction
In 95 DEG C of heating 2-5min in PCR instrument, make the abundant sex change of template DNA, then enter amplification cycles, 94 DEG C of 30sec, 53-58 DEG C of 40sec, 72 DEG C of 35sec, carry out 27-30 circulation, finally, keep 5min, 4-16 DEG C of preservation at 72 DEG C;
(4) agarose gel electrophoresis detects
After pcr amplification reaction terminates, get product and carry out agarose gel electrophoresis detection, and observe with ultraviolet gel imaging system, take a picture, contrast according to electrophoresis result and the standard diagram provided, if there is no band or only have a band, then judge that this DNA sample is from Fujian-East Guangdong race large yellow croaker.
3. the discrimination method of Dai-ju stock Pseudosciaena crocea according to claim 2 and Fujian-East Guangdong race large yellow croaker, it is characterized in that large yellow croaker template DNA 10ng in the pcr amplification reaction system of primer pair J1, large yellow croaker DNA40ng in the pcr amplification reaction system of primer pair J2, PCR response procedures is: 95 DEG C of 3min, amplification cycles 94 DEG C of 30sec, 55.8 DEG C of 40sec, 72 DEG C of 35sec totally 28 circulations, 72 DEG C of 5min, 4 DEG C of preservations.
4. the discrimination method of Dai-ju stock Pseudosciaena crocea according to claim 3 and Fujian-East Guangdong race large yellow croaker, it is characterized in that PCR reaction terminate after, get product 10ul with 1.5% agarose gel electrophoresis detection.
5. the discrimination method of the Dai-ju stock Pseudosciaena crocea according to claim 3 or 4 and Fujian-East Guangdong race large yellow croaker, is characterized in that adopting phenol chloroform extraction method to extract large yellow croaker DNA.
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Non-Patent Citations (5)
| Title |
|---|
| 大黄鱼线粒体DNA控制区遗传多样性分析;毛勇等;《厦门大学学报(自然科学版)》;20100530;第49卷(第03期);440-444 * |
| 大黄鱼线粒体DNA的限制性内切酶图谱;李明云等;《科技通报》;20060825;第22卷(第04期);456-461 * |
| 岱衢族大黄鱼（Pseudosciaena crocea）线粒体基因组全序列的扩增及其特异鉴别引物的开发;沈锡权等;《海洋与湖沼》;20130515;第44卷(第3期);摘要部分，表3，图2，3讨论部分 * |
| 舟山小黄鱼线粒体DNA D-loop区序列变异的遗传多样性分析;郑文娟等;《动物学研究》;20120608;第33卷(第3期);329-336 * |
| 闽-粤东族与岱衢族养殖大黄鱼的遗传多样性研究;黄良敏等;《厦门大学学报(自然科学版)》;20061130;第45卷(第06期);836-839 * |
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