CN101366944A - Mrpink在抑制罗氏沼虾精子活性中的应用 - Google Patents
Mrpink在抑制罗氏沼虾精子活性中的应用 Download PDFInfo
- Publication number
- CN101366944A CN101366944A CNA2007100708087A CN200710070808A CN101366944A CN 101366944 A CN101366944 A CN 101366944A CN A2007100708087 A CNA2007100708087 A CN A2007100708087A CN 200710070808 A CN200710070808 A CN 200710070808A CN 101366944 A CN101366944 A CN 101366944A
- Authority
- CN
- China
- Prior art keywords
- mrpink
- rosenbergii
- spermatozoon
- germ cell
- macrobracium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 230000000452 restraining effect Effects 0.000 title 1
- 230000000694 effects Effects 0.000 claims abstract description 17
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 claims abstract description 11
- 210000004995 male reproductive system Anatomy 0.000 claims abstract description 11
- 239000000137 peptide hydrolase inhibitor Substances 0.000 claims abstract description 11
- 241001327110 Macrobrachium rosenbergii Species 0.000 claims description 18
- 239000002299 complementary DNA Substances 0.000 claims description 16
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 5
- 239000003112 inhibitor Substances 0.000 claims description 4
- 241000238557 Decapoda Species 0.000 abstract description 9
- 238000011160 research Methods 0.000 abstract description 6
- 230000002401 inhibitory effect Effects 0.000 abstract description 3
- 210000004602 germ cell Anatomy 0.000 abstract 7
- 239000013505 freshwater Substances 0.000 abstract 3
- 108090000623 proteins and genes Proteins 0.000 description 17
- 108010010803 Gelatin Proteins 0.000 description 5
- 229920000159 gelatin Polymers 0.000 description 5
- 239000008273 gelatin Substances 0.000 description 5
- 235000019322 gelatine Nutrition 0.000 description 5
- 235000011852 gelatine desserts Nutrition 0.000 description 5
- 239000003708 ampul Substances 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 239000007952 growth promoter Substances 0.000 description 3
- 230000013632 homeostatic process Effects 0.000 description 3
- 108020004999 messenger RNA Proteins 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000238559 Macrobrachium Species 0.000 description 2
- 229940122344 Peptidase inhibitor Drugs 0.000 description 2
- 108090000708 Proteasome Endopeptidase Complex Proteins 0.000 description 2
- 108091036066 Three prime untranslated region Proteins 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 238000001976 enzyme digestion Methods 0.000 description 2
- 210000000981 epithelium Anatomy 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 238000012214 genetic breeding Methods 0.000 description 2
- 125000001475 halogen functional group Chemical group 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 210000004994 reproductive system Anatomy 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 210000002955 secretory cell Anatomy 0.000 description 2
- 230000002194 synthesizing effect Effects 0.000 description 2
- 108091026890 Coding region Proteins 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 108091026898 Leader sequence (mRNA) Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 238000000636 Northern blotting Methods 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 102000004245 Proteasome Endopeptidase Complex Human genes 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 108091034057 RNA (poly(A)) Proteins 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000005138 cryopreservation Methods 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000002468 fat body Anatomy 0.000 description 1
- 210000004996 female reproductive system Anatomy 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 210000002816 gill Anatomy 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 210000003677 hemocyte Anatomy 0.000 description 1
- 229940000351 hemocyte Drugs 0.000 description 1
- 210000000514 hepatopancreas Anatomy 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 238000007901 in situ hybridization Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 230000001418 larval effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 230000020509 sex determination Effects 0.000 description 1
- 235000015170 shellfish Nutrition 0.000 description 1
- 230000009303 sperm storage Effects 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明提供了罗氏沼虾雄性生殖系统特异性Kazal型蛋白酶抑制MRPINK在抑制罗氏沼虾精子活性中的应用。在研究中发现，在罗氏沼虾的精子中添加MRPINK能有效的抑制精子活性，使精卵不能受精。在精子的冻存中添加类似的蛋白酶抑制剂，抑制精子内酶活性，使精子不活化，对精子的低温储存具有重要的借鉴意义。
Description
(一)技术领域
本发明涉及罗氏沼虾雄性生殖系统特异性Kazal型蛋白酶抑制剂MRPINK在抑制罗氏沼虾精子活性中的应用。
(二)背景技术
生物的性别、生殖现象是生物学研究的主要内容之一，它涉及到许多重大的生物学问题，比如性别决定、遗传、发育和进化等，因此历来倍受关注。目前对于性别、生殖的分子机制研究主要集中在人类和模式生物中，而对于一般水生生物这方面的研究很少，尤其在甲壳类中，已知的相关分子信息更加缺乏。
虾类作为具有重要经济价值的水生生物，在分子水平的研究开展得相对较少，因此，大量获取水生动物性别和生殖相关基因资源来研究它们的生殖生物学、遗传育种等显得尤为必要。虾类同龄雌雄性别个体的大小、生长速度相差悬殊，性别与生殖制约着它们的产值。若能培育出单性虾，并进行单性化养殖可以大幅度提高上市商品规格和养殖产量，因此虾类性别控制一直是遗传育种研究的热点之一。目前有关这方面的认识仅仅局限在形态结构和一般的组织化学、生化成份和基本的生理指标，在基因水平的认识和可操作性则几乎为零。
(三)发明内容
本发明则是通过对罗氏沼虾雄性生殖相关基因进行克隆、表达特征分析以及定位功能等研究，提供了罗氏沼虾雄性生殖系统特异性Kazal型蛋白酶抑制剂MRPINK在抑制罗氏沼虾精子活性中的应用。
本发明采用的技术方案是：
罗氏沼虾雄性生殖系统特异性Kazal型蛋白酶抑制剂MRPINK在抑制罗氏沼虾精子活性中的应用。(有关MRPINK的相关介绍参见文献：Cao JX，Dai JQ，Dai ZM，Yin GL and Yang WJ.A male-reproduction-relatedkazal-type peptidase inhibitor gene in the prawn，Macrobrachium rosenbergii：molecular characterization and expression patterns.Marine Biotechnology.2007，9：45～55.)
所述MRPINK的cDNA序列如下：AGTTGACAGACTGCCAGCTTCCTGTGCAGAATTCTGAACGATAGGCTTAGGTAAGACCTCACAACTAGCAAAATGAAACTGTACATCTTCACCTTCTGCCTAGCCGCGTCAATTCTCGTCGGGTTTTCATCTGGGGAATACCGCTTCCTTGTCTCTCCACTGTGCCCCGTACGTTGTACTCTGAGGTACATCCCTGTCTGTGGCAGCGACGGCCGAACGTACGGGAACAAATGTCACCTGGACAATGCACGTCTCTGTGACAACAGCAATGTTCAGGTAGTTCACGAAGGCGAATGTAAGCAAGACTACTGCGCCCCGGTAGTCCGCTGCCCACCGGTCATAAAAGCCGTATGTGCCAACGACGGCCAAACATACTTGAACGAGTGTTTCGCCAAAGTCGCAGCCTGCGGATTCCCTAACCTGAAGATAGTCCACAGCGGATTATGTGGTCCTAGACCAAAGACTGAAGTATGGTAGCGAGGTGGTGACTGTTGGAAATAGACCATTACGTGAGTGAAAACGGCGATATATGAATATGTTCCATCAATCAAATCAAGTTCTGATCGGCTGTGTACTAATAACTTCGTACATATGGCCTTCTTTCAAGTTCTCCTGATTTCCACCAGATTTATTATATTTTTATGGATATTGGAGGTTTCTGAGATACGCCCAGTTGTCTATTACATCTGAAGAAAAACTTGATCAATAAAGCCTTTGTAACAGGAAAAAAAAAAAA。
具体的，所述应用为：以MRPINK制备精子活性抑制剂，用于罗氏沼虾精子冻存。
优选的，所述精子活性抑制剂为MRPINK。所述MRPINK的氨基酸序列如下：MKLYIFTFCLAASILVGFSSGEYRFLVSPLCPVRCTLRYIPVCGSDGRTYGNKCHLDNARLCDNSNVQVVHEGECKQDYCAPVVRCPPVIKAVCANDGQTYLNECFAKVAACGFPNLKIVHSGLCGPRPKTEVW。
选取罗氏沼虾几乎全组织，包括肌肉、肝胰腺、心脏、鳃、胃、血细胞、雄性生殖系统、卵巢、脑、中枢神经、脂肪体、眼柄、肠和腹部神经进行Northernblotting分析，结果表明MRPINK基因仅表达在雄性生殖系统，而且在输精管中的表达占主导地位，其次是末端壶腹，在精巢中的表达并不是很高。
通过分析表明MRPINK基因mRNA的表达在后幼体期四周内检测不到，此后其表达出现，但表达量随着罗氏沼虾逐渐生长发育并无明显变化。MRPINK基因的表达在罗氏沼虾的不同生长发育时期呈现恒定趋势，表明这一基因更多的与维持内环境稳定和整个组织的完整性相关，而不是调控精子形成或形态分化的特异阶段。
运用原位杂交的实验手段在细胞水平上检测了MRPINK在罗氏沼虾雄性生殖系统中的定位，发现这MRPINK基因的mRNA仅表达在输精管和末端壶腹的内壁上皮分泌细胞中。这一特异性的定位暗示着它可能在精子自身保护、生殖系统中蛋白酶降解及内环境稳定等过程中发挥功能。
MRPINK基因的表达在罗氏沼虾生长发育时期呈现恒定趋势且仅在输精管和末端壶腹的内壁上皮分泌细胞中有表达，表明这一基因在精子自身保护、生殖系统中蛋白酶降解及维持内环境稳定等过程中发挥了重要功能。MRPINK是蛋白酶抑制剂，参与维持雄性生殖系统内环境的稳定，对精子的生理成熟和活力维持可能有十分重要的意义。在我们的研究中发现，在罗氏沼虾的精子中添加MRPINK能有效的抑制精子活性。
人类精子库已应用于临床治疗，但是作为生育能力的储备形式仍然有很大的局限性。目前人类精子储存的办法是将精子冻存在装有液态氮的大罐子中，在超低温情况下保持精子自身能量不被消耗。但在精子冷冻和复苏过程中，温度的变化会对精子产生生物物理损伤和化学损害，使精子细胞膜水、电介质浓缩、紊乱、使不稳固的类脂蛋白变性、以及机械性破坏从而影响精子的生命活力。此外，精子冷冻的价格非常昂贵，需要一套严格的控制系统，以避免出现差错。
在精子的冻存中添加类似的蛋白酶抑制剂，抑制精子内酶活性，使精子不活化，对精子的低温储存具有重要的借鉴意义。
(四)具体实施方式
下面结合具体实施例对本发明进行进一步描述，但本发明的保护范围并不仅限于此：
实施例1：MRPINK基因及其编码的蛋白质的获得
具体可参见文献：Cao JX，Dai JQ，Dai ZM，Yin GL and Yang WJ.Amale-reproduction-related kazal-type peptidase inhibitor gene in the prawn，Macrobrachium rosenbergii：molecular characterization and expressionpatterns.Marine Biotechnology.2007，9：45～55.
①取罗氏沼虾雄性和雌性生殖系统组织，按Trizol试剂盒说明抽提总RNA；
②建立罗氏沼虾雄性生殖系统SSH cDNA文库，筛选差异性表达的基因片段：
A.第一条链的合成：按Oligotex mRNA Kits法，提取PolyA+RNA，以PolyA+RNA为模板反转录合成单链cDNA。
B.双链(ds cDNA)的合成，Rsa I酶消化产生平末端。
C.接头连接：把Rsa I酶切后雄性组织ds cDNA分成两份，分别加上接头1(Adaptor1)和接头2(Adaptor2)，16℃连接过夜，形成两组检测子(Tester)。
D.消减杂交：
第一，分别将Tester1和Tester2的连接产物与RsaI酶消化的过量驱动子(Driver)ds cDNA混合，在98℃下热变性，然后在65～68℃下放置10小时使Tester和Driver的cDNA进行充分地杂交。检测子单链cDNA片段被均化，即平均高、低丰度的cDNA浓度，使其基本相等。
第二，将第一次杂交的产物混匀到一起，然后在68℃继续放置过夜(约16小时)，使Tester1和Tester2之间的目的差异cDNA形成异源杂交子。
E.巢氏PCR扩增差异表达的cDNA。
F.筛选差异性文库：PCR产物经连接转化到TOPO10载体上，构建差异表达基因富集的cDNA文库，筛选差异性表达的基因。
③全长cDNA的获得：
利用FirstChoiceTM RLM-RACE kit合成5’和3’RACE，分别得到487bp的5’端和270bp的3’端，拼接完成后获得全长cDNA，命名为MRPINK基因。
MRPINK cDNA序列如下：
AGTTGACAGACTGCCAGCTTCCTGTGCAGAATTCTGAACGATAGGC
TTAGGTAAGACCTCACAACTAGCAAAATGAAACTGTACATCTTCAC
CTTCTGCCTAGCCGCGTCAATTCTCGTCGGGTTTTCATCTGGGGAAT
ACCGCTTCCTTGTCTCTCCACTGTGCCCCGTACGTTGTACTCTGAGG
TACATCCCTGTCTGTGGCAGCGACGGCCGAACGTACGGGAACAAA
TGTCACCTGGACAATGCACGTCTCTGTGACAACAGCAATGTTCAGG
TAGTTCACGAAGGCGAATGTAAGCAAGACTACTGCGCCCCGGTAG
TCCGCTGCCCACCGGTCATAAAAGCCGTATGTGCCAACGACGGCCA
AACATACTTGAACGAGTGTTTCGCCAAAGTCGCAGCCTGCGGATTC
CCTAACCTGAAGATAGTCCACAGCGGATTATGTGGTCCTAGACCAA
AGACTGAAGTATGGTAGCGAGGTGGTGACTGTTGGAAATAGACCA
TTACGTGAGTGAAAACGGCGATATATGAATATGTTCCATCAATCAA
ATCAAGTTCTGATCGGCTGTGTACTAATAACTTCGTACATATGGCC
TTCTTTCAAGTTCTCCTGATTTCCACCAGATTTATTATATTTTTATGG
ATATTGGAGGTTTCTGAGATACGCCCAGTTGTCTATTACATCTGAA
GAAAAACTTGATCAATAAAGCCTTTGTAACAGGAAAAAAAAAAAA
RPINK cDNA全长736bp，编码区405bp，5’和3’非翻译区长度分别是72bp和247bp，其中3’非翻译区具有多聚腺苷酸信号。该基因已递交到GenBank，其序列号为AY885242。
MRPINK氨基酸序列如下：
MKLYIFTFCLAASILVGFSSGEYRFLVSPLCPVRCTLRYIPVCGSDGRTY
GNKCHLDNARLCDNSNVQVVHEGECKQDYCAPVVRCPPVIKAVCAN
DGQTYLNECFAKVAACGFPNLKIVHSGLCGPRPKTEVW。
它编码135个氨基酸，分子量约14.7kDa，推演的氨基酸序列中具有两个Kazal型domain，且含有21个氨基酸的信号肽。
MRPINK和已知的Kazal型蛋白酶抑制剂在氨基酸序列上的同源性为16～32％，其Kazal型domain序列上有19～52％的同源性，表明这个蛋白属于Kazal家族中的成员。MRPINK作为蛋白酶抑制剂，与已知哺乳动物蛋白酶抑制剂可能具有相似的功能。
实施例2：MRPINK对罗氏沼虾的精子活性的抑制作用
明胶底物实验：
罗氏沼虾精子(挑选成熟雄性罗氏沼虾，摘取壶腹磨碎，离心，计数，使精子浓度为1×106cells/ml)用MRPINK(2mg/ml)28℃预处理1h；未经MRPINK处理的罗氏沼虾精子作为对照组；将处理后样品涂于已含明胶(终浓度为5％)的玻片上，37℃孵育过夜，相差显微镜(400×)下观察。
实验结果表明：95％罗氏沼虾精子会降解明胶形成晕轮(图1-A)，经MRPINK处理的精子不能降解明胶形成晕轮(图1-B)，MRPINK能有效抑制罗氏沼虾精子降解明胶的活性。由此说明，MRPINK对罗氏沼虾的精子活性有抑制作用。
序列表_ST25
SEQUENCE LISTING
<110>浙江大学
<120>MRPINK在抑制罗氏沼虾精子活性中的应用
<130>
<160>2
<170>PatentIn version 3.4
<210>1
<211>736
<212>DNA
<213>Macrobrachium rosenbergii
<400>1
<210>2
<211>134
<212>PRT
<213>Macrobrachium rosenbergii
<400>2
Claims (5)
1.罗氏沼虾雄性生殖系统特异性Kazal型蛋白酶抑制剂MRPINK在抑制罗氏沼虾精子活性中的应用。
2.如权利要求1所述的应用，其特征在于所述MRPINK的cDNA序列如下
AGTTGACAGACTGCCAGCTTCCTGTGCAGAATTCTGAACGATAG
GCTTAGGTAAGACCTCACAACTAGCAAAATGAAACTGTACATCT
TCACCTTCTGCCTAGCCGCGTCAATTCTCGTCGGGTTTTCATCTG
GGGAATACCGCTTCCTTGTCTCTCCACTGTGCCCCGTACGTTGTA
CTCTGAGGTACATCCCTGTCTGTGGCAGCGACGGCCGAACGTAC
GGGAACAAATGTCACCTGGACAATGCACGTCTCTGTGACAACAG
CAATGTTCAGGTAGTTCACGAAGGCGAATGTAAGCAAGACTACT
GCGCCCCGGTAGTCCGCTGCCCACCGGTCATAAAAGCCGTATGT
GCCAACGACGGCCAAACATACTTGAACGAGTGTTTCGCCAAAGT
CGCAGCCTGCGGATTCCCTAACCTGAAGATAGTCCACAGCGGAT
TATGTGGTCCTAGACCAAAGACTGAAGTATGGTAGCGAGGTGGT
GACTGTTGGAAATAGACCATTACGTGAGTGAAAACGGCGATATA
TGAATATGTTCCATCAATCAAATCAAGTTCTGATCGGCTGTGTAC
TAATAACTTCGTACATATGGCCTTCTTTCAAGTTCTCCTGATTTCC
ACCAGATTTATTATATTTTTATGGATATTGGAGGTTTCTGAGATA
CGCCCAGTTGTCTATTACATCTGAAGAAAAACTTGATCAATAAA
GCCTTTGTAACAGGAAAAAAAAAAAA。
3.如权利要求2所述的应用，其特征在于所述应用为：以MRPINK制备精子活性抑制剂，用于罗氏沼虾精子冻存。
4.如权利要求3所述的应用，其特征在于所述精子活性抑制剂为MRPINK。
5.如权利要求1～4之一所述的应用，其特征在于所述MRPINK的氨基酸序列如下：
MKLYIFTFCLAASILVGFSSGEYRFLVSPLCPVRCTLRYIPVCGSDGRT
YGNKCHLDNARLCDNSNVQVVHEGECKQDYCAPVVRCPPVIKAVC
ANDGQTYLNECFAKVAACGFPNLKIVHSGLCGPRPKTEVW。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100708087A CN101366944B (zh) | 2007-08-16 | 2007-08-16 | Mrpink在抑制罗氏沼虾精子活性中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100708087A CN101366944B (zh) | 2007-08-16 | 2007-08-16 | Mrpink在抑制罗氏沼虾精子活性中的应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101366944A true CN101366944A (zh) | 2009-02-18 |
| CN101366944B CN101366944B (zh) | 2011-05-11 |
Family
ID=40411036
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2007100708087A Expired - Fee Related CN101366944B (zh) | 2007-08-16 | 2007-08-16 | Mrpink在抑制罗氏沼虾精子活性中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101366944B (zh) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104745584A (zh) * | 2015-04-12 | 2015-07-01 | 浙江万里学院 | 一种用于干扰罗氏沼虾雄性生殖的核酸分子及其制备方法 |
-
2007
- 2007-08-16 CN CN2007100708087A patent/CN101366944B/zh not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104745584A (zh) * | 2015-04-12 | 2015-07-01 | 浙江万里学院 | 一种用于干扰罗氏沼虾雄性生殖的核酸分子及其制备方法 |
| CN104745584B (zh) * | 2015-04-12 | 2017-07-04 | 浙江万里学院 | 一种用于干扰罗氏沼虾雄性生殖的核酸分子及其制备方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101366944B (zh) | 2011-05-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Grasso et al. | Microarray analysis identifies candidate genes for key roles in coral development | |
| Yeong Kwon et al. | Molecular characterization of putative yolk processing enzymes and their expression during oogenesis and embryogenesis in rainbow trout (Oncorhynchus mykiss) | |
| Nie et al. | The HSP70 gene expression responses to thermal and salinity stress in wild and cultivated Manila clam Ruditapes philippinarum | |
| MacLean et al. | Rhox: a new homeobox gene cluster | |
| Meng et al. | Inducible variation in anaerobic energy metabolism reflects hypoxia tolerance across the intertidal and subtidal distribution of the Pacific oyster (Crassostrea gigas) | |
| CN110747279B (zh) | 暗纹东方鲀snp分子标记及其在遗传育种的应用 | |
| Larsen et al. | Intraspecific variation in expression of candidate genes for osmoregulation, heme biosynthesis and stress resistance suggests local adaptation in European flounder (Platichthys flesus) | |
| Zhang et al. | Transcriptome analysis of five ovarian stages reveals gonad maturation in female Macrobrachium nipponense | |
| Tward et al. | Identification of the alternative oxidase gene and its expression in the copepod Tigriopus californicus | |
| Segev-Hadar et al. | Genome editing using the CRISPR-Cas9 system to generate a solid-red germline of Nile tilapia (Oreochromis niloticus) | |
| Tepolt et al. | Rapid adaptation to temperature via a potential genomic island of divergence in the invasive green crab, Carcinus maenas | |
| Callaghan et al. | Expression of sex and reproduction-related genes in Marsupenaeus japonicus | |
| CN101148668B (zh) | 猪产肉性状相关基因btg1的克隆及其在猪分子标记辅助选择中的应用 | |
| CN106119407A (zh) | Lap3基因作为绵羊生长性状的分子标记及其应用 | |
| Kong et al. | Cloning, expression and localization of the Daphnia carinata transformer gene DcarTra during different reproductive stages | |
| Yoshida et al. | Establishment of enhancer detection lines expressing GFP in the gut of the ascidian Ciona intestinalis | |
| CN101366944B (zh) | Mrpink在抑制罗氏沼虾精子活性中的应用 | |
| Liu et al. | Discovery of Nanos1 and Nanos2/3 as germ cell markers during scallop gonadal development | |
| Lee et al. | Assessment of suitable reference genes for rt-qpcr normalization with developmental samples in pacific abalone haliotis discus hannai | |
| Cheng et al. | Research trends of development on pearl bivalve mollusks based on a bibliometric network analysis in the past 25 years | |
| Zhu et al. | Cardiac performance and heart gene network provide dynamic responses of bay scallop Argopecten irradians irradians exposure to marine heatwaves | |
| Ding et al. | Transcriptome analysis of blood for the discovery of sex-related genes in ricefield eel Monopterus albus | |
| Li et al. | Integrative biological analyses of responses to food deprivation reveal resilience mechanisms in sea urchin larvae | |
| Cucini et al. | First de novo transcriptome analysis of the Antarctic springtail Cryptopygus terranovus (Collembola: Isotomidae) following mid-term heat exposure | |
| Ichida et al. | Characterization of a vasa homolog in Mekong giant catfish (Pangasianodon gigas): Potential use as a germ cell marker |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20110511 Termination date: 20140816 |
|
| EXPY | Termination of patent right or utility model |