CN102021227A - Method for positioning target gene of pinctada fucata martenssi - Google Patents
Method for positioning target gene of pinctada fucata martenssi Download PDFInfo
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- CN102021227A CN102021227A CN2009100932081A CN200910093208A CN102021227A CN 102021227 A CN102021227 A CN 102021227A CN 2009100932081 A CN2009100932081 A CN 2009100932081A CN 200910093208 A CN200910093208 A CN 200910093208A CN 102021227 A CN102021227 A CN 102021227A
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- methyl alcohol
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- pinctada fucata
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- 241000490568 Pinctada fucata Species 0.000 title claims abstract description 42
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 30
- 238000000034 method Methods 0.000 title claims abstract description 26
- 238000007901 in situ hybridization Methods 0.000 claims abstract description 15
- 239000000523 sample Substances 0.000 claims abstract description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 111
- 239000000243 solution Substances 0.000 claims description 48
- 229930040373 Paraformaldehyde Natural products 0.000 claims description 35
- 229920002866 paraformaldehyde Polymers 0.000 claims description 35
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 claims description 33
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 16
- 210000001161 mammalian embryo Anatomy 0.000 claims description 15
- 239000000203 mixture Substances 0.000 claims description 15
- 239000002904 solvent Substances 0.000 claims description 15
- 238000013507 mapping Methods 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 8
- 239000011780 sodium chloride Substances 0.000 claims description 8
- 238000005406 washing Methods 0.000 claims description 8
- 229920001213 Polysorbate 20 Polymers 0.000 claims description 7
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 7
- 108010006777 nacrein Proteins 0.000 claims description 7
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 7
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 5
- 239000007864 aqueous solution Substances 0.000 claims description 4
- 239000011049 pearl Substances 0.000 abstract description 9
- 230000009456 molecular mechanism Effects 0.000 abstract description 4
- 238000011160 research Methods 0.000 abstract description 4
- 230000013020 embryo development Effects 0.000 abstract 1
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- -1 Methane amide Chemical class 0.000 description 4
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- 238000006073 displacement reaction Methods 0.000 description 4
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- 210000002966 serum Anatomy 0.000 description 4
- 238000010186 staining Methods 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000000903 blocking effect Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 241000237519 Bivalvia Species 0.000 description 2
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- SHIBSTMRCDJXLN-UHFFFAOYSA-N Digoxigenin Natural products C1CC(C2C(C3(C)CCC(O)CC3CC2)CC2O)(O)C2(C)C1C1=CC(=O)OC1 SHIBSTMRCDJXLN-UHFFFAOYSA-N 0.000 description 1
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- HLFSDGLLUJUHTE-SNVBAGLBSA-N Levamisole Chemical compound C1([C@H]2CN3CCSC3=N2)=CC=CC=C1 HLFSDGLLUJUHTE-SNVBAGLBSA-N 0.000 description 1
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Abstract
The invention discloses a method for positioning a target gene in pinctada fucata martenssi and provides a method for positioning the target gene of the pinctada fucata martenssi in the embryonic development process. The method comprises the following steps of: performing whole-embryo in-situ hybridization on the pinctada fucata martenssi; and positioning the target gene according to the result of the whole-embryo in-situ hybridization, wherein a probe used for the whole-embryo in-situ hybridization is a probe for the target gene. A gene positioning method is provided for the research of the pinctada fucata martenssi and better technology is provided for further research of various molecular mechanisms of the pinctada fucata martenssi. The method is easy to operate, does not need special equipment, has low cost and plays an important role in improving the quality of pearl shells and pearls.
Description
Technical field
The present invention relates to a kind of method of the goal gene in the pinctada fucata being carried out the assignment of genes gene mapping.
Background technology
Pearl is a kind of typical biomineralization product, and it is not only the starting material of ornament and medicine, makeup, but also is design and the desirable template for preparing novel bionical high-performance engineering material.Therefore the molecular mechanism of studying the pearl mineralising has important theory and practice significance.
Pinctada fucata (Pinctada fucata Martenssi), claim pteria martensii again, belong to Mollusca (Mollusca), lamellibranchiata (Bivalvia), Margarita order (Pterioida), Pteriidae (Pteriidae), Pinctada (Pinctada), mainly being distributed in China and Japanese coastal, is to be used for the main shellfish that sea water pearls is produced at present in the world.
At present, pearl mineralising Study on Mechanism is mainly concentrated on albumen or the separation of polypeptide and the clone of gene thereof who participates in mineralising, also there are many difficulties in the functional study of related gene.Study the location of functional gene, will help to disclose the function of gene, illustrate the molecular mechanism of nacre mineralising, thereby be that output and the quality that improves pearl provided fundamental basis.
Visitor in the invention
The purpose of this invention is to provide a kind of method of the goal gene in the pinctada fucata being carried out the assignment of genes gene mapping.
The method that goal gene in the pinctada fucata is carried out the assignment of genes gene mapping provided by the invention is that pinctada fucata is carried out full embryo in situ hybridization, according to full embryo results of in situ hybridization goal gene is carried out the assignment of genes gene mapping; The used probe of described full embryo in situ hybridization is the probe at goal gene.
Described full embryo in situ hybridization can comprise the steps:
1) pinctada fucata is fixed;
2) pinctada fucata that step 1) is obtained carries out again aquation, shells and fixes;
3) with step 2) pinctada fucata carry out prehybridization;
4) pinctada fucata of step 3) is hybridized;
5) pinctada fucata with step 4) detects.
Described step 1) can adopt following steps: with pinctada fucata with 4 ℃ of 4% paraformaldehyde solutions fixedly more than the 24h; Use 50% methyl alcohol-2% paraformaldehyde solution and methanol wash then successively;
Described 4% paraformaldehyde solution, solute are Paraformaldehyde 96, and solvent is water or PBS, and the final concentration of Paraformaldehyde 96 is 4% (quality percentage composition); Described 50% methyl alcohol-2% paraformaldehyde solution, solute is methyl alcohol and Paraformaldehyde 96, and solvent is a water, and the final concentration of methyl alcohol is 50% (quality percentage composition), and the final concentration of Paraformaldehyde 96 is 2% (quality percentage composition); The preparation method of described PBS is as follows: NaCl 8g, KCl 0.2g, Na
2HPO
41.44g, KH
2PO
40.24g, H
2O 1L, adjust pH to 7.4.
The time of described usefulness 50% methyl alcohol-2% paraformaldehyde solution washing specifically can be 5 minutes; The described time with methanol wash specifically can be 5 minutes.
Described step 2) can adopt following steps: the pinctada fucata that step 1) obtains is used the PBST solution of 50% methyl alcohol, the PBST solution aquation of 30% methyl alcohol successively, with the PBST washing, repeat once then; EDTA with 0.2mol handles then, and the PBS solution that is replaced as 4% Paraformaldehyde 96 is again fixed, and washes twice with PBST at last;
The PBST solution of described 50% methyl alcohol, solute are methyl alcohol, and solvent is PBST, and the final concentration of methyl alcohol is 50% (quality percentage composition); The PBST solution of described 30% methyl alcohol, solute are methyl alcohol, and solvent is PBST, and the final concentration of methyl alcohol is 30% (quality percentage composition); Described PBST preparation method is as follows: adding Tween-20 in PBS, to make its final concentration be 0.1% (quality percentage composition); The PBS solution of described 4% Paraformaldehyde 96, solute are Paraformaldehyde 96, and solvent is PBS, and the final concentration of Paraformaldehyde 96 is 4% (quality percentage composition); Described PBS preparation method is as follows: NaCl 8g, KCl 0.2g, Na
2HPO
41.44g, KH
2PO
40.24g, H
2O 1L, adjust pH to 7.4.
Described step 2) specifically can adopt following steps: the pinctada fucata that step 1) is obtained is used the PBST solution aquation 5 minutes of PBST solution, 30% methyl alcohol of 50% methyl alcohol successively, with PBST washing 5 minutes, repeats once then; Handle half an hour with the EDTA aqueous solution of 0.2mol/L then, the PBS solution that is replaced as 4% Paraformaldehyde 96 is again fixed 20 minutes, washes twice with PBST at last, each 5 minutes.
More than in arbitrary described method, described goal gene can be the nacrein gene.The sequence of described nacrein gene specifically can be as GENBANK ACCESS ION NUMBER D83523.In the described full embryo in situ hybridization, used probe specifically can be shown in the sequence 1 of sequence table.Described pinctada fucata specifically can be the pinctada fucata of setting stage.
The present invention provides the method for the assignment of genes gene mapping for the research of pinctada fucata, for the further various molecular mechanisms of research pinctada fucata provide good technology.Method easy handling of the present invention need not specific installation, and cost is lower.The present invention is significant to the quality of the quality of Margarita and pearl.
Description of drawings
Fig. 1 is the full embryo in situ hybridization result of nacrein in the setting stage.
Embodiment
Following embodiment is convenient to understand better the present invention, but does not limit the present invention.Experimental technique among the following embodiment if no special instructions, is ordinary method.Used test materials among the following embodiment if no special instructions, is to buy from routine biochemistry reagent shop and obtains.Following % all refers to the quality percentage composition if no special instructions.
The prescription of used solution is as follows among the embodiment:
One, base soln prescription
20×SSC：
Na3Citrate 2H
2O 88.2g, NaCl 175.5g, DEPC H
2O to 1L; Suction filtration, sterilization.
Two, the prescription of used solution among the embodiment
(1)PBS：
NaCl 8g, KCl 0.2g, Na
2HPO
41.44g, KH
2PO
40.24g, DEPC H
2O 1L; HCl adjust pH to 7.4, suction filtration, sterilization.
(2) 4% Paraformaldehyde 96s:
Paraformaldehyde 96 40g, PBS 1L; The heating and continuous solution that is stirred to is clarified;-20 ℃ of preservations.
(3)PBST：
Adding Tween-20 among the PBS, to make its final concentration be 0.1%.
(4)2×SSCT：
10 times of 20 * SSC dilutions add Tween-20, and making its final concentration is 0.2%.
(5)0.2×SSCT
10 times of 2 * SSCT dilutions.
(6)HYB-：
Methane amide: 20 * SSC liquid storage: DEPC water=2: 1: 1 preparation, adding Tween-20 then, to make its final concentration be 0.1% ,-20 ℃ of preservations.
(7)HYB+：
HYB-20ml, yeast RNA 10mg, heparin 1mg;-20 ℃ of preservations.
(8)MAB：
Maleic acid 11.6g, NaCl 8.8g; Transferring to the pH value with solid NaOH (about 7g) is 7.5,4 ℃ of preservations.
(9)MABT：
Adding Tween-20 among the MAB, to make its final concentration be 0.1%.
(10)10％blocking?solution：
Blocking reagent (Luo Shi 11096176001) 8g, MAB 72ml.
(11) 1: 2: 7 solution:
Deactivation sheep blood serum: 10%blocking reagent: MABT=1: 2: 7 (volume ratio); Time spent now joins.
(12)Staining?buffer：
Tris 12.1g, Mgcl 6H
2O 10.2g, NaCl 5.85g, Tween-20 1ml, sterilized water 1L; Transfer pH value to 9.5.
Embodiment, the nacrein gene in the pinctada fucata is carried out the assignment of genes gene mapping
The pinctada fucata (pearl farm, military camp, Guangxi) of setting stage is carried out full embryo in situ hybridization, and concrete steps are as follows:
1, preparation probe
Extract the cDNA of pinctada fucata, carry out RT-PCR, obtain purpose fragment (DNA shown in the sequence 1), use digoxigenin labeled after enzyme is cut.
2, the collection of young shellfish is with fixing:
The pinctada fucata of setting stage is fixed with 4% Paraformaldehyde 96,, guaranteed that the set time is more than 24h in 4 ℃ of preservations.(solute is methyl alcohol and Paraformaldehyde 96, and solvent is a water with 50% methyl alcohol-2% paraformaldehyde solution behind the 24h; In this solution, the final concentration of methyl alcohol is 50%, and the final concentration of Paraformaldehyde 96 is 2%) wash, room temperature is placed 5min, after change 100% methyl alcohol into, room temperature is placed 5min, is replaced as 100% methyl alcohol again, and is in-20 ℃ of preservations, stand-by.
3, aquation is again shelled and is fixed
(solute is a methyl alcohol, and solvent is PBST, and the final concentration of methyl alcohol is 50% to use the PBST solution of 50%, 30% methyl alcohol successively; Solute is a methyl alcohol, and solvent is PBST, and the final concentration of methyl alcohol is 30%) aquation 5 minutes, be replaced as PBST solution, placed 5 minutes, repeat once; The EDTA aqueous solution with 0.2mol/L is handled half an hour, is replaced as 4% Paraformaldehyde 96 and fixes 20 minutes, washes twice with PBST, places room temperature five minutes at every turn.
4, prehybridization
Be replaced as about 300ul HYB-solution in each pipe, vibration is avoided in 60 ℃ of water-baths 5 minutes.Replace HYB-solution with isopyknic HYB+ solution, 60 ℃ of water-baths, prehybridization is more than 4 hours.
5, hybridization
1) inhales the HYB+ that removes prehybridization, add that 100ul has added the HYB+ solution (concentration and probe concentration is about 1ng/ul) of sex change (75 ℃ of sex change 10min) probe.
2) 60 ℃ of temperature are bathed and are spent the night.
6, detect
1) probe is reclaimed, and use 1 milliliter of 50% methane amide/2 * SSCT solution (solute is a methane amide, and solvent is 2 * SSCT, and the final concentration of methane amide is 50%) successively, 1 milliliter of 2 * SSCT, 1 milliliter of 0.2 * SSCT washing.
2) wash twice with MABT, each five minutes, be placed on shaking table and shake gently; Add 1: 2: 7 solution of 1ml under the room temperature, the time is one hour; Add enzyme in 1: 3000 ratio in the solution at 1: 2: 7 and connect DigiTAb (Luo Shi; Article No. 1745832), 4 ℃ of refrigerator overnight.
3) contain MABT solution (solute is hot inactivated serum, and solvent is MABT, and the final concentration of hot inactivated serum is 10%) the displacement antibody-solutions (the recyclable utilization of antibody repeatedly) of 10% hot inactivated serum with 1ml, place shaking table last 25 minute; Then with 1ml MABT displacement, 25 minutes; Again with the displacement of 1ml MABT solution, more than one hour; At last with the displacement of 1ml MABT solution, 25 minutes.
4) with 1ml contain the left-handed rice of 1mM azoles Staining buffer (with before in Staining buffer, add the LEVAMISOLE HCL liquid storage of 1M, making it final concentration is 1mM) wash twice, placed five minutes at every turn.
5) embryo is changed in 96 orifice plates, inhale and to remove staining buffer, add 180ul BM Purple APSubstrate (substrate, with before add left-handed meter azoles of 5mM) (Luo Shi; Article No. 11745832910), masking foil is wrapped with lucifuge in 40 octal plates outside, avoids shaking 28.5 ℃ of incubator colour developings.
6) observe the embryo every 20 minutes and whether begin colour developing.
7) will develop the color substrate sucking-off among the embryo completely adds that 4% is fixing after washing twice or thrice with PBST, takes pictures.4 ℃ of refrigerators are preserved.
The results are shown in Figure 1.What as seen from Figure 1, nacrein was special expresses at mantle.
Sequence table
＜110〉Tsing-Hua University
＜120〉localization method of pinctada fucata goal gene
<130>CGGNARY92539
<160>1
<210>1
<211>501
<212>DNA
＜213〉artificial sequence
<220>
<223>
<400>1
cgattgggcc?cgacgtcgca?tgctcccggc?cgccatggcg?gccgcgggaa?ttcgattgga 60
tcgcccttgt?gaaggtcctc?attattggca?aaccatatca?caatgcttca?ttgcgtgtgg 120
aattggacag?agacaatctc?caatcaacat?cgtttcttat?gatgctaaat?ttcttcagcg 180
tttgccaaag?ttgaaattca?agccacatat?ggagaaatta?aaaacagaag?tgaccaatca 240
tcagaaccga?gctccagagt?tcgagccaga?ggatggggaa?aatctgtacg?tgaagctaaa 300
taacctagtg?gacggtcatt?ataaattcca?taatcttcac?gttcataatg?gtagaaccgg 360
acgtaaggga?tcagaacaca?gtgttaacgg?tcgtttcaca?cctatggagg?ctcatttggt 420
tttccatcat?gatgaacaaa?cacactttga?acctacacgc?acaagggcga?tcccaatcac 480
tagtgaattc?gcggccgcct?g 501
Claims (10)
1. the method that the goal gene in the pinctada fucata is carried out the assignment of genes gene mapping is that pinctada fucata is carried out full embryo in situ hybridization, according to full embryo results of in situ hybridization goal gene is carried out the assignment of genes gene mapping; The used probe of described full embryo in situ hybridization is the probe at goal gene.
2. the method for claim 1, it is characterized in that: described full embryo in situ hybridization comprises the steps:
1) pinctada fucata is fixed;
2) pinctada fucata that step 1) is obtained carries out again aquation, shells and fixes;
3) with step 2) pinctada fucata carry out prehybridization;
4) pinctada fucata of step 3) is hybridized;
5) pinctada fucata with step 4) detects.
3. method as claimed in claim 2 is characterized in that: in the described step 1): with pinctada fucata with 4 ℃ of 4% paraformaldehyde solutions fixedly more than the 24h; Use 50% methyl alcohol-2% paraformaldehyde solution and methanol wash then successively;
Described 4% paraformaldehyde solution, solute are Paraformaldehyde 96, and solvent is water or PBS, and the final concentration of Paraformaldehyde 96 is 4% (quality percentage composition); Described 50% methyl alcohol-2% paraformaldehyde solution, solute is methyl alcohol and Paraformaldehyde 96, and solvent is a water, and the final concentration of methyl alcohol is 50% (quality percentage composition), and the final concentration of Paraformaldehyde 96 is 2% (quality percentage composition); The preparation method of described PBS is as follows: NaCl 8g, KCl 0.2g, Na
2HPO
41.44g, KH
2PO
40.24g, H
2O 1L, adjust pH to 7.4.
4. method as claimed in claim 3 is characterized in that: the time of described usefulness 50% methyl alcohol-2% paraformaldehyde solution washing is 5 minutes; The described time with methanol wash is 5 minutes.
5. as arbitrary described method in the claim 2 to 4, it is characterized in that: described step 2): the pinctada fucata that step 1) obtains is used the PBST solution of 50% methyl alcohol, the PBST solution aquation of 30% methyl alcohol successively, with the PBST washing, repeat once then; Handle with the EDTA aqueous solution of 0.2M then, the PBS solution that is replaced as 4% Paraformaldehyde 96 is again fixed, and washes twice with PBST at last;
The PBST solution of described 50% methyl alcohol, solute are methyl alcohol, and solvent is PBST, and the final concentration of methyl alcohol is 50% (quality percentage composition); The PBST solution of described 30% methyl alcohol, solute are methyl alcohol, and solvent is PBST, and the final concentration of methyl alcohol is 30% (quality percentage composition); The preparation method of described PBST is as follows: adding Tween-20 in PBS, to make its final concentration be 0.1% (quality percentage composition); The PBS solution of described 4% Paraformaldehyde 96, solute are Paraformaldehyde 96, and solvent is PBS, and the final concentration of Paraformaldehyde 96 is 4% (quality percentage composition); The preparation method of described PBS is as follows: NaCl 8g, KCl 0.2g, Na
2HPO
41.44g, KH
2PO
40.24g, H
2O 1L, adjust pH to 7.4.
6. method as claimed in claim 5, it is characterized in that: described step 2): the pinctada fucata that step 1) is obtained is used the PBST solution aquation 5 minutes of the PBST solution of described 50% methyl alcohol, described 30% methyl alcohol successively, with described PBST washing 5 minutes, repeat once then; Handle half an hour with the EDTA aqueous solution of described 0.2M then, the PBS solution that is replaced as described 4% Paraformaldehyde 96 is again fixed 20 minutes, washes twice with described PBST at last, each 5 minutes.
7. as arbitrary described method in the claim 1 to 6, it is characterized in that: described goal gene is the nacrein gene.
8. method as claimed in claim 8 is characterized in that: described nacrein gene is the DNA shown in the GENBANK ACCESSIONNUMBER D83523.
9. method as claimed in claim 8 is characterized in that: the used probe of described full embryo in situ hybridization is shown in the sequence 1 of sequence table.
10. as arbitrary described method in the claim 1 to 9, it is characterized in that: described pinctada fucata is the pinctada fucata of setting stage.
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|---|---|---|---|
| CN2009100932081A CN102021227A (en) | 2009-09-15 | 2009-09-15 | Method for positioning target gene of pinctada fucata martenssi |
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|---|---|---|---|
| CN2009100932081A CN102021227A (en) | 2009-09-15 | 2009-09-15 | Method for positioning target gene of pinctada fucata martenssi |
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|---|---|
| CN102021227A true CN102021227A (en) | 2011-04-20 |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103535303A (en) * | 2013-10-24 | 2014-01-29 | 厦门大学 | Method for fixing gastropod mollusk |
| CN104099415A (en) * | 2014-06-24 | 2014-10-15 | 中国科学院南海海洋研究所 | Detection primer marked by SNP and associated with pinctada martensi adductor muscle weight and application of detection primer |
| CN104561305A (en) * | 2014-12-31 | 2015-04-29 | 中国科学院南海海洋研究所 | SNP298299 marker significantly correlated with pinctada martensii mollusc part weight and adductor muscle weight, as well as primers and application of SNP298299 marker |
| CN104711343A (en) * | 2014-12-31 | 2015-06-17 | 中国科学院南海海洋研究所 | SNP411871 marker associated to shell mould and weight of pinctada martensii, primer and application thereof |
| CN104911188A (en) * | 2015-04-28 | 2015-09-16 | 中国水产科学研究院南海水产研究所 | Pinctada fucata epidermal growth factor acceptor gene (Pf-EGFR) and application thereof |
| CN109207612A (en) * | 2018-11-15 | 2019-01-15 | 中国水产科学研究院黄海水产研究所 | Detection method for overall in-situ hybridization of marine medusa scyphistoma |
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2009
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103535303A (en) * | 2013-10-24 | 2014-01-29 | 厦门大学 | Method for fixing gastropod mollusk |
| CN103535303B (en) * | 2013-10-24 | 2015-06-10 | 厦门大学 | Method for fixing gastropod mollusk |
| CN104099415A (en) * | 2014-06-24 | 2014-10-15 | 中国科学院南海海洋研究所 | Detection primer marked by SNP and associated with pinctada martensi adductor muscle weight and application of detection primer |
| CN104099415B (en) * | 2014-06-24 | 2015-09-23 | 中国科学院南海海洋研究所 | A kind of detection primer of the SNP marker associated with pteria martensii closed shell flesh heavy phase and application thereof |
| CN104561305A (en) * | 2014-12-31 | 2015-04-29 | 中国科学院南海海洋研究所 | SNP298299 marker significantly correlated with pinctada martensii mollusc part weight and adductor muscle weight, as well as primers and application of SNP298299 marker |
| CN104711343A (en) * | 2014-12-31 | 2015-06-17 | 中国科学院南海海洋研究所 | SNP411871 marker associated to shell mould and weight of pinctada martensii, primer and application thereof |
| CN104911188A (en) * | 2015-04-28 | 2015-09-16 | 中国水产科学研究院南海水产研究所 | Pinctada fucata epidermal growth factor acceptor gene (Pf-EGFR) and application thereof |
| CN109207612A (en) * | 2018-11-15 | 2019-01-15 | 中国水产科学研究院黄海水产研究所 | Detection method for overall in-situ hybridization of marine medusa scyphistoma |
| CN109207612B (en) * | 2018-11-15 | 2021-08-31 | 中国水产科学研究院黄海水产研究所 | Detection method for overall in-situ hybridization of marine medusa scyphistoma |
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