CN103667327B - 一种用于检测环境污染物的转基因斑马鱼的制备方法 - Google Patents
一种用于检测环境污染物的转基因斑马鱼的制备方法 Download PDFInfo
- Publication number
- CN103667327B CN103667327B CN201310654584.XA CN201310654584A CN103667327B CN 103667327 B CN103667327 B CN 103667327B CN 201310654584 A CN201310654584 A CN 201310654584A CN 103667327 B CN103667327 B CN 103667327B
- Authority
- CN
- China
- Prior art keywords
- zebrafish
- gfp
- transgenic
- expression vector
- sequence
- 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.)
- Active
Links
- 241000252212 Danio rerio Species 0.000 title claims abstract description 52
- 230000009261 transgenic effect Effects 0.000 title claims abstract description 28
- 238000002360 preparation method Methods 0.000 title claims description 5
- 238000012360 testing method Methods 0.000 title claims description 3
- 101100400865 Mus musculus Abcb1b gene Proteins 0.000 claims abstract description 12
- 239000013604 expression vector Substances 0.000 claims abstract description 10
- 108020004414 DNA Proteins 0.000 claims abstract description 7
- 238000011144 upstream manufacturing Methods 0.000 claims abstract description 6
- 230000001568 sexual effect Effects 0.000 claims abstract description 4
- 230000002068 genetic effect Effects 0.000 claims abstract description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 5
- 210000001161 mammalian embryo Anatomy 0.000 claims description 4
- 108091081024 Start codon Proteins 0.000 claims description 3
- 238000002347 injection Methods 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- 238000000520 microinjection Methods 0.000 claims description 2
- 238000013467 fragmentation Methods 0.000 claims 2
- 238000006062 fragmentation reaction Methods 0.000 claims 2
- 239000003344 environmental pollutant Substances 0.000 abstract description 21
- 210000002257 embryonic structure Anatomy 0.000 abstract description 13
- 231100000719 pollutant Toxicity 0.000 abstract description 12
- 239000000126 substance Substances 0.000 abstract description 12
- 230000014509 gene expression Effects 0.000 abstract description 9
- 229910001385 heavy metal Inorganic materials 0.000 abstract description 8
- 239000012634 fragment Substances 0.000 abstract description 7
- 238000000034 method Methods 0.000 abstract description 7
- 125000005575 polycyclic aromatic hydrocarbon group Chemical group 0.000 abstract description 7
- 235000013601 eggs Nutrition 0.000 abstract description 5
- 230000001105 regulatory effect Effects 0.000 abstract description 4
- 108091026890 Coding region Proteins 0.000 abstract description 3
- 102000034287 fluorescent proteins Human genes 0.000 abstract description 3
- 108091006047 fluorescent proteins Proteins 0.000 abstract description 3
- 238000010367 cloning Methods 0.000 abstract description 2
- 238000010276 construction Methods 0.000 abstract description 2
- 238000012258 culturing Methods 0.000 abstract description 2
- 230000013011 mating Effects 0.000 abstract description 2
- 230000001419 dependent effect Effects 0.000 abstract 1
- 239000005090 green fluorescent protein Substances 0.000 description 16
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 11
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 10
- YKYOUMDCQGMQQO-UHFFFAOYSA-L cadmium dichloride Chemical compound Cl[Cd]Cl YKYOUMDCQGMQQO-UHFFFAOYSA-L 0.000 description 6
- 230000007613 environmental effect Effects 0.000 description 5
- 108700024526 zebrafish sox32 Proteins 0.000 description 5
- 238000001514 detection method Methods 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 231100000419 toxicity Toxicity 0.000 description 4
- 230000001988 toxicity Effects 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 229940011871 estrogen Drugs 0.000 description 3
- 239000000262 estrogen Substances 0.000 description 3
- 238000012544 monitoring process Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000003053 toxin Substances 0.000 description 3
- 231100000765 toxin Toxicity 0.000 description 3
- 108700012359 toxins Proteins 0.000 description 3
- 239000013598 vector Substances 0.000 description 3
- 102000018832 Cytochromes Human genes 0.000 description 2
- 108010052832 Cytochromes Proteins 0.000 description 2
- 241000282414 Homo sapiens Species 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- 231100000517 death Toxicity 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000004720 fertilization Effects 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 150000003071 polychlorinated biphenyls Chemical class 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- 108010006533 ATP-Binding Cassette Transporters Proteins 0.000 description 1
- 102000005416 ATP-Binding Cassette Transporters Human genes 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- VWDXGKUTGQJJHJ-UHFFFAOYSA-N Catenarin Natural products C1=C(O)C=C2C(=O)C3=C(O)C(C)=CC(O)=C3C(=O)C2=C1O VWDXGKUTGQJJHJ-UHFFFAOYSA-N 0.000 description 1
- 241000193901 Dreissena polymorpha Species 0.000 description 1
- 239000010282 Emodin Substances 0.000 description 1
- RBLJKYCRSCQLRP-UHFFFAOYSA-N Emodin-dianthron Natural products O=C1C2=CC(C)=CC(O)=C2C(=O)C2=C1CC(=O)C=C2O RBLJKYCRSCQLRP-UHFFFAOYSA-N 0.000 description 1
- 101150066002 GFP gene Proteins 0.000 description 1
- YOOXNSPYGCZLAX-UHFFFAOYSA-N Helminthosporin Natural products C1=CC(O)=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O YOOXNSPYGCZLAX-UHFFFAOYSA-N 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- NTGIIKCGBNGQAR-UHFFFAOYSA-N Rheoemodin Natural products C1=C(O)C=C2C(=O)C3=CC(O)=CC(O)=C3C(=O)C2=C1O NTGIIKCGBNGQAR-UHFFFAOYSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 229910052793 cadmium Inorganic materials 0.000 description 1
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 description 1
- WLZRMCYVCSSEQC-UHFFFAOYSA-N cadmium(2+) Chemical compound [Cd+2] WLZRMCYVCSSEQC-UHFFFAOYSA-N 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- RHMXXJGYXNZAPX-UHFFFAOYSA-N emodin Chemical compound C1=C(O)C=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O RHMXXJGYXNZAPX-UHFFFAOYSA-N 0.000 description 1
- VASFLQKDXBAWEL-UHFFFAOYSA-N emodin Natural products OC1=C(OC2=C(C=CC(=C2C1=O)O)O)C1=CC=C(C=C1)O VASFLQKDXBAWEL-UHFFFAOYSA-N 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 231100000613 environmental toxicology Toxicity 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- PKUBGLYEOAJPEG-UHFFFAOYSA-N physcion Natural products C1=C(C)C=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O PKUBGLYEOAJPEG-UHFFFAOYSA-N 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 108010054624 red fluorescent protein Proteins 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 150000001911 terphenyls Chemical class 0.000 description 1
- 231100000167 toxic agent Toxicity 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
本发明公开了一种用于检测环境污染物的转基因斑马鱼的制备方法，包括：(1)从斑马鱼基因组中克隆Mdr1蛋白的启动子调控序列；(2)将克隆的DNA片段融合于GFP编码序列的上游构建表达载体；(3)将该表达载体进行斑马鱼胚胎显微注射；(4)将注射后的胚胎培养至性成熟，并与野生型斑马鱼交配产卵；(5)荧光显微镜下筛选阳性的F1代，继续自交传至F2代，得到稳定遗传的转基因品系。由于荧光蛋白的表达水平可受到重金属及多环芳烃等多种环境污染物浓度依赖性的诱导，因而本案所制备的转基因斑马鱼可用于探测环境中的上述化学污染物的存在及浓度，且具有经济、快速、方便、准确的优点。
Description
技术领域
本发明涉及一种环境污染物生物探测器的生产方法，具体涉及一种灵敏监测环境污染物浓度的转基因斑马鱼的构建方法。
背景技术
随着社会的发展，人类生产和利用的合成化学物质越来越多，这些物质通过各种途径进入环境中，不仅会引起水生生物死亡和水生态系统破坏，还会直接或间接威胁到人类健康。据世界卫生组织估计，重金属、多环芳烃及多氯联苯等化学污染物导致世界每年大约数百万人死亡，更是60～90％癌症的诱因。仅在中国，每年因工业污染物引发的新增癌症患者数量就高达94.5万。然而，当今社会已无法离开化学工业及其产品，为此，通过加强对环境中污染物水平的检测，进而限制工厂污染物排放就变得更为紧迫。
作为一种来自印度的热带淡水鱼，斑马鱼最初被用于发育生物学与遗传学的研究。由于其具有自身的独特优势，如：体型小、体外受精、卵生且产卵量大、胚胎透明且发育速度快，此外其培养液中可直接加入化学污染物，进而检测污染物毒性。因此，斑马鱼也逐渐被应用于环境毒理学研究中，而欧盟、美国及日本各国更是将斑马鱼列入环境保护标准测试生物中。
与此同时，根据污染物作用于斑马鱼后的生物学指标变化，结合绿色荧光蛋白(GFP)及红色荧光蛋白等转基因技术，研究者还构建了各种转基因斑马鱼模型使其可用于定量反映环境污染物浓度。如根据多环芳烃对于细胞色素酶(CYP)1A1的诱导作用，Hung等人构建了表达绿色荧光蛋白(CYP-GFP)的转基因斑马鱼，可在0.04μg/L多氯化三联苯的诱导下表达绿色荧光，提示环境中多环芳烃的含量超标(Hung K.W.，Suen M.F.，Chen Y.F.，Cai H.B.，MoZ.X.，Yung K.K.；Detection of water toxicity using cytochrome P450transgenic zebrafish as live biosensor：for polychlorinated biphenylstoxicity；Biosens Bioelectron，2012，31(1)，548-553)。复旦大学宋后燕、钟涛领衔的课题组则筛选出对雌激素类物质敏感的斑马鱼vtg基因启动子，据此构建了随vtg表达绿色荧光蛋白进而可直观监测环境中雌激素含量的转基因斑马鱼，响应浓度可达到μg/L级(Chen H.，Hu J.，Yang J.，WangY.，Xu H.，Jiang Q.，Gong Y.，Gu Y.，Song H.，Generation of a fluorescenttransgenic zebrafish for detection of environmental estrogens.AquatToxicol，2010，96(1)，53-61)。与传统环境检测方法相比，转基因斑马鱼模型具有快捷方便且经济有效等优势，但目前已有的监测模型往往仅能针对特定化学污染物进行反应，缺乏广谱性。
作为机体抵御外源物摄入的主要工具之一，Mdr1等ABC转运蛋白被发现高表达于受精24h后的斑马鱼中，可显著降低重金属毒素、多环芳烃和多氯联苯等多种毒素的积累及其毒性(Faria M.，Navarro A.，Luckenbach T.，Pina B.，Barata C.，Characterization of the multixenobiotic resistance(MXR)mechanism in embryos and larvae of the zebra mussel(Dfeissenapolymorpha)and studies on its role in tolerance to single and mixturecombinations of toxicants.AquatToxicol，2011，101(1)，78-87)。此外，我们的实验结果及文献(He Q.，Liu K.，Wang S.，Hou H.，Yuan Y.，WangX.，Toxicity induced by emodin on zebrafish embryos.Drug Chem Toxicol，2012，35(2)，149-154)均表明，Mdr1可在毒素存在的情况下被诱导表达，从而提示可针对性地开发新的环境监测型斑马鱼。
发明内容
本发明的目的是提供一种用于检测环境污染物的转基因斑马鱼的制备方法，该转基因斑马鱼可方便快捷地对多种化学污染物浓度进行监测，为水环境保护工作提供新的工具。
为实现上述技术目的，达到上述技术效果，本发明通过以下技术方案实现：
一种用于检测环境污染物的转基因斑马鱼的制备方法，包括以下步骤：
步骤(1)，从斑马鱼基因组中克隆Mdr1蛋白的启动子调控序列；
步骤(2)，将步骤(1)中克隆的DNA片段融合于GFP编码序列的上游，构建成表达载体Mdr1promoter-GFP；
步骤(3)，将所述表达载体Mdr1promoter-GFP进行斑马鱼胚胎显微注射；
步骤(4)，将注射后的胚胎培养至性成熟，并与野生型斑马鱼交配产卵；
步骤(5)，荧光显微镜下筛选阳性的F1代，继续自交传至F2代，得到稳定遗传的转基因品系。
优选地，所述的用于检测环境污染物的转基因斑马鱼的制备方法，步骤(1)中克隆的DNA片段来源于Mdr1蛋白编码基因的ATG起始密码子5’端上游的280bp序列。
优选地，所述的用于检测环境污染物的转基因斑马鱼的制备方法，在步骤(3)中，所述表达载体Mdr1promoter-GFP注入的时间点为斑马鱼受精卵单细胞时期。
本发明的有益效果是：通过本案的制备方法生产出的转基因斑马鱼可稳定表达绿色荧光蛋白，且蛋白表达水平受外源性化学污染物浓度依赖性的诱导，因而该转基因斑马鱼可用于探测环境中的重金属及多环芳烃等化学污染物的存在及浓度，且其具有经济、快速、方便、准确等优点。
附图说明
图1为转基因斑马鱼胚胎培养至24hpf时，氯化镉处5h后，GFP基因表达被氯化镉浓度依赖性的诱导的PCR结果图。
具体实施方式
下面结合附图对本发明做进一步的详细说明，以令本领域技术人员参照说明书文字能够据以实施。
实施例1-转基因斑马鱼的制备方法
(1)从斑马鱼基因组中克隆Mdr1蛋白的启动子调控序列，即Mdr1蛋白编码基因的ATG起始密码子的5’端上游280bp的一段DNA序列；
通过mRNA序列与斑马鱼基因组序列的BLASTIN程序比对分析，在GENEBANK上找到斑马鱼Mdr1基因的启动子序列，设计对应的正反向引物，两端分别接入XhoI和BamHI的酶切稳点，高保真PCR的方法扩增到片段，此片段进一步连接到pGME-T载体中，经过测序后确认此克隆为目标片段。
(2)构建表达载体mdr1promoter-GFP
用XhoI和BamHI将斑马鱼Mdr1启动子280bp从载体上切下，装入pEGFP载体中，所得到的重组表达载体mdr1promoter-GFP，包含斑马鱼Mdr1基因的调控序列和GFP编码序列。
(3)显微注射
收集单细胞时期的斑马鱼受精卵，将约2μl的载体溶液装入微电极中，显微注射到细胞浆中，每个胚胎约2nl，胚胎培养到性成熟。
(4)转基因斑马鱼的获得
将转基因斑马鱼与野生型杂交，通过重金属镉处理和荧光筛选，获得具有稳定绿色荧光表达的转基因斑马鱼。之后提取胚胎的基因组DNA，用GFP的特异性引物来扩增出对应大小的片段进行测序确认。
实施例2一转基因斑马鱼环境监测效果验证
(1)配制含一系列浓度氯化镉(Cd)的斑马鱼培养液
分别配制含10ug/L、100ug/L、1000ug/L Cd的斑马鱼培养液以及不含Cd的空白斑马鱼培养液。
(2)重金属Cd诱导斑马鱼荧光蛋白的表达
24hpf预培养后，斑马鱼胚胎置于含系列浓度Cd的培养液中，5hpf后，荧光显微镜下镜检，可验证胚胎随Cd浓度的升高，表达出更强的绿色荧光。其他重金属亦可诱导斑马鱼荧光蛋白的表达。
(3)PCR验证
提取经Cd预处理后的胚胎DNA，PCR检测其中GFP的表达情况，请参见图1，可知，随Cd浓度的提高，GFP表达量逐渐升高。即该转基因斑马鱼可良好地检测水环境中镉离子的存在。在本实施例中只列举了Cd的数据，该转基因斑马鱼同样可检测水环境中其他重金属离子及多环芳烃的存在。
尽管本发明的实施方案已公开如上，但其并不仅仅限于说明书和实施方式中所列运用，它完全可以被适用于各种适合本发明的领域，对于熟悉本领域的人员而言，可容易地实现另外的修改，因此在不背离权利要求及等同范围所限定的一般概念下，本发明并不限于特定的细节和这里示出与描述的图例。
Claims (1)
1.一种用于检测环境污染物的转基因斑马鱼的制备方法，其特征在于，包括以下步骤：
步骤(1)，从斑马鱼基因组中克隆Mdr1蛋白的启动子调控序列；
步骤(2)，将步骤(1)中克隆的DNA片段融合于GFP编码序列的上游，构建成表达载体Mdr1promoter-GFP；
步骤(3)，将所述表达载体Mdr1promoter-GFP进行斑马鱼胚胎显微注射；
步骤(4)，将注射后的胚胎培养至性成熟，并与野生型斑马鱼交配产卵；
步骤(5)，荧光显微镜下筛选阳性的F1代，继续自交传至F2代，得到稳定遗传的转基因品系；
步骤(1)中克隆的DNA片段来源于Mdr1蛋白编码基因的ATG起始密码子5’端上游的280bp序列；
在步骤(3)中，所述表达载体Mdr1promoter-GFP注入的时间点为斑马鱼受精卵单细胞时期。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310654584.XA CN103667327B (zh) | 2013-12-06 | 2013-12-06 | 一种用于检测环境污染物的转基因斑马鱼的制备方法 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310654584.XA CN103667327B (zh) | 2013-12-06 | 2013-12-06 | 一种用于检测环境污染物的转基因斑马鱼的制备方法 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103667327A CN103667327A (zh) | 2014-03-26 |
| CN103667327B true CN103667327B (zh) | 2015-10-21 |
Family
ID=50306096
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310654584.XA Active CN103667327B (zh) | 2013-12-06 | 2013-12-06 | 一种用于检测环境污染物的转基因斑马鱼的制备方法 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103667327B (zh) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104542389B (zh) * | 2014-12-23 | 2017-01-25 | 中国科学院苏州生物医学工程技术研究所 | 一种非酒精性脂肪肝斑马鱼的制备方法 |
| CN106070063B (zh) * | 2016-07-07 | 2019-03-05 | 贵州医科大学 | 带abcb4的转基因斑马鱼系及其建立方法 |
| CN106282359A (zh) * | 2016-08-31 | 2017-01-04 | 中国科学院重庆绿色智能技术研究院 | 一种利用整体原位杂交技术检测环境污染物发育毒性的方法 |
| CN106777893A (zh) * | 2016-11-23 | 2017-05-31 | 大连理工大学 | 一种利用可移动污染物探测器辨识城市空间多污染物源的方法 |
| CN107460240B (zh) * | 2017-08-03 | 2018-12-07 | 广东省实验动物监测所 | 一种利用基因表达变化快速检测海水或海洋沉积物中多环芳烃的方法 |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1083788B1 (en) * | 1998-05-31 | 2005-08-10 | The University of Georgia Research Foundation, Inc. | Transgenic fish carrying a bacteriophage-derived transgene construct for mutation detection |
| CN102023199A (zh) * | 2009-09-11 | 2011-04-20 | 华东理工大学 | 一种通过转基因斑马鱼胚胎模型测试纳米材料毒性的方法 |
| CN102539640A (zh) * | 2010-12-17 | 2012-07-04 | 中国科学院生态环境研究中心 | 一种鱼类早期生命阶段的高通量急性毒性测试方法 |
| CN102899384A (zh) * | 2012-11-13 | 2013-01-30 | 中国科学院广州生物医药与健康研究院 | 应用斑马鱼测试有机溶剂毒性的方法 |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040147030A1 (en) * | 2000-05-22 | 2004-07-29 | Nebert Daniel W | Transgenic animals for monitoring water quality |
-
2013
- 2013-12-06 CN CN201310654584.XA patent/CN103667327B/zh active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1083788B1 (en) * | 1998-05-31 | 2005-08-10 | The University of Georgia Research Foundation, Inc. | Transgenic fish carrying a bacteriophage-derived transgene construct for mutation detection |
| CN102023199A (zh) * | 2009-09-11 | 2011-04-20 | 华东理工大学 | 一种通过转基因斑马鱼胚胎模型测试纳米材料毒性的方法 |
| CN102539640A (zh) * | 2010-12-17 | 2012-07-04 | 中国科学院生态环境研究中心 | 一种鱼类早期生命阶段的高通量急性毒性测试方法 |
| CN102899384A (zh) * | 2012-11-13 | 2013-01-30 | 中国科学院广州生物医药与健康研究院 | 应用斑马鱼测试有机溶剂毒性的方法 |
Non-Patent Citations (2)
| Title |
|---|
| Hung K.W.et al.Detection of water toxicity using cytochrome P450 transgenic zebrafish as live biosensor:for polychlorinated biphenyls toxicity.《Biosensor and Bioelectronics》.2012,第31卷(第1期),第548-553页. * |
| Toxicity induced by emodin on zebrafish embryos;QIUXIA HE et al;《Drug Chem Toxicol》;20120430;第35卷(第2期);第149-154页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103667327A (zh) | 2014-03-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Kong et al. | PHB3 maintains root stem cell niche identity through ROS-responsive AP2/ERF transcription factors in Arabidopsis | |
| Kim et al. | Recruitment of rod photoreceptors from short-wavelength-sensitive cones during the evolution of nocturnal vision in mammals | |
| CN103667327B (zh) | 一种用于检测环境污染物的转基因斑马鱼的制备方法 | |
| Portmann et al. | Behavioral abnormalities and circuit defects in the basal ganglia of a mouse model of 16p11. 2 deletion syndrome | |
| Firnhaber et al. | Neuron-specific feeding RNAi in C. elegans and its use in a screen for essential genes required for GABA neuron function | |
| Yamamoto et al. | Functional and evolutionary diversification of Otx2 and Crx in vertebrate retinal photoreceptor and bipolar cell development | |
| Yu et al. | A P-loop NTPase regulates quiescent center cell division and distal stem cell identity through the regulation of ROS homeostasis in Arabidopsis root | |
| Houwing et al. | A role for Piwi and piRNAs in germ cell maintenance and transposon silencing in Zebrafish | |
| Hiller et al. | Hundreds of conserved non-coding genomic regions are independently lost in mammals | |
| Wen et al. | Adaptive regulation of testis gene expression and control of male fertility by the Drosophila hairpin RNA pathway | |
| Abe et al. | The origin of the bifurcated axial skeletal system in the twin-tail goldfish | |
| Ten Hove et al. | SCHIZORIZA encodes a nuclear factor regulating asymmetry of stem cell divisions in the Arabidopsis root | |
| Liu et al. | Changes throughout a genetic network mask the contribution of Hox gene evolution | |
| Brito et al. | Transcriptomic and functional analyses of the piRNA pathway in the Chagas disease vector Rhodnius prolixus | |
| Shirayama et al. | The Vasa Homolog RDE-12 engages target mRNA and multiple argonaute proteins to promote RNAi in C. elegans | |
| Langevin et al. | FTR83, a member of the large fish-specific finTRIM family, triggers IFN pathway and counters viral infection | |
| Mao et al. | Characterization of RAG1 and IgM (mu chain) marking development of the immune system in red-spotted grouper (Epinephelus akaara) | |
| López-Cuadros et al. | Isolation of the sex-determining gene Sex-lethal (Sxl) in Penaeus (Litopenaeus) vannamei (Boone, 1931) and characterization of its embryogenic, gametogenic, and tissue-specific expression | |
| Zou et al. | A conserved role of αA-crystallin in the development of the zebrafish embryonic lens | |
| Bergeron et al. | Brain selective transgene expression in zebrafish using an NRSE derived motif | |
| England et al. | Hmx3a has essential functions in zebrafish spinal cord, ear and lateral line development | |
| Thermes et al. | Medaka simplet (FAM53B) belongs to a family of novel vertebrate genes controlling cell proliferation | |
| Voogdt et al. | Evolutionary regression and species-specific codon usage of TLR15 | |
| Nhnhkorn et al. | Penaeus monodon IKKs participate in regulation of cytokine-like system and antiviral responses of innate immune system | |
| van Gelderen et al. | miR-210 promotes immune-and suppresses oocyte meiosis-related genes in the zebrafish ovarian cells |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | 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 |