斑马鱼中囊胚转换分子机制的研究
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摘要
中囊胚转换(Midblastula transition，MBT)在胚胎发育过程中是一个重要的过程。从这个时期开始表达的基因将在后续的原肠胚形成及体节形成过程中起至关重要的作用。存MBT前，早期胚胎发育主要受母型mRNAs和蛋白质调控，转录水平较低。以前依靠突变的方法发现了许多重要的基因是与发育相关的。但是，还有很多存早期发育过程中具有重要功能的基因没有被发现，因为这些基因的突变会引起胚胎死亡。
本实验应用美国Affymetrix公司的斑马鱼基因表达谱芯片，对斑马鱼中囊胚转换过程中的基因表达差异进行了研究。中囊胚转换大致发生在胚胎发育至2.75hpf(hour post-fertilization)。我们分别提取发育至2.5hpf和4hpf的斑马鱼胚胎总RNA。芯片分析结果表明，共获得了479个差异表达的基因，其中360个基因表达上调，119个基因表达下调。通过GO(Gene Ontology)分析表明，上调表达的基因按功能可以分为以下几种类别：信号转导，细胞通讯，表达调控，转录因子活性，胚胎发育，形态发生，性别分化，代谢以及转运类蛋白等。其中可能有一些与中囊胚转换相关并具有重要作用的基因，但其具体功能尚不清楚。选取表达上调率较高，且在中囊胚转换前未表达的31个基因作为进一步研究对
activity is regulated by CK2 phosphorylation of cyclin H. Oncogene. 2002,21 (33):5031-5037.
24 Wu L, Chen P, Shum C H, et al. MAT 1-modulated CAK activity regulates cell cycle G(l) exit. Mol Cell Biol. 2001,21 (l):260-270.
25 Kato J Y, Matsuoka M, Strom D K, et al. Regulation of cyclin D-dependent kinase 4 (cdk4) by cdk4-activating kinase. Mol Cell Biol. 1994,14 (4) :2713-2721.
26 Gerber M R, Farrell A, Deshaies R J, et al. Cdc37 is required for association of the protein kinase Cdc28 with Gl andmito.tic cyclins. Proc Natl Acad Sci USA. 1995,92 (10):4651-4655.
27 He Q, Peng H, Collins S J, et al. Retinoid-modulated MAT1 ubiquitination and CAK activity. Faseb J. 2004,18 (14): 1734-1736.
28 Jordan P, Cunha C, Carmo-Fonseca M. The cdk7-cyclin H-MAT1 complex associated with TFIIH is localized in coiled bodies. Mol Biol Cell. 1997,8 (7):1-207-1217.
29 Tassan J P, Schultz S J, Bartek J, et al. Cell cycle analysis of the activity, subcellular localization, and subunit composition of human CAK (CDK-activating kinase). J Cell Biol. 1994,127 (2):467-478.
30 Lamond A 1, Earnshaw W C. Structure and function in the nucleus. Science. 1998,280 (5363):547-553.
31 Faust M, Kartarius S, Schwindling S L, et al. Cyclin H is a new binding partner for protein kinase CK2. Biochem Biophys Res Commun. 2002,296 (1): 13-19.
32 Gannon J V, Nebreda A, Goodger N M, et al. A measure of the mitotic index: studies of the abundance and half-life of p34cdc2 in cultured cells and normal and neoplastic tissues. Genes Cells. 1998,3 (l):17-27.
33 Kim J M, McGaughy J T, Bogle R K, et al. Meiotic expression of the cyclin H/Cdk7 complex in male germ cells of the mouse. Biol Reprod. 2001,64 (5): 1400-1408.
34 Kaldis P, Solomon M J. Analysis of CAK activities from human cells. Eur J Biochem. 2000,267 (13):4213-4221.
35 Leclerc V, Raisin S, Leopold P. Dominant-negative mutants reveal a role for the Cdk7 kinase at the mid-blastula transition in Drosophila embryos. Embo J. 2000,19 (7):1567-1575.
36 Hermand D, Pihlak A, Westerling T, et al. Fission yeast Cskl is a
本实验应用美国Affymetrix公司的斑马鱼基因表达谱芯片，对斑马鱼中囊胚转换过程中的基因表达差异进行了研究。中囊胚转换大致发生在胚胎发育至2.75hpf(hour post-fertilization)。我们分别提取发育至2.5hpf和4hpf的斑马鱼胚胎总RNA。芯片分析结果表明，共获得了479个差异表达的基因，其中360个基因表达上调，119个基因表达下调。通过GO(Gene Ontology)分析表明，上调表达的基因按功能可以分为以下几种类别：信号转导，细胞通讯，表达调控，转录因子活性，胚胎发育，形态发生，性别分化，代谢以及转运类蛋白等。其中可能有一些与中囊胚转换相关并具有重要作用的基因，但其具体功能尚不清楚。选取表达上调率较高，且在中囊胚转换前未表达的31个基因作为进一步研究对
activity is regulated by CK2 phosphorylation of cyclin H. Oncogene. 2002,21 (33):5031-5037.
24 Wu L, Chen P, Shum C H, et al. MAT 1-modulated CAK activity regulates cell cycle G(l) exit. Mol Cell Biol. 2001,21 (l):260-270.
25 Kato J Y, Matsuoka M, Strom D K, et al. Regulation of cyclin D-dependent kinase 4 (cdk4) by cdk4-activating kinase. Mol Cell Biol. 1994,14 (4) :2713-2721.
26 Gerber M R, Farrell A, Deshaies R J, et al. Cdc37 is required for association of the protein kinase Cdc28 with Gl andmito.tic cyclins. Proc Natl Acad Sci USA. 1995,92 (10):4651-4655.
27 He Q, Peng H, Collins S J, et al. Retinoid-modulated MAT1 ubiquitination and CAK activity. Faseb J. 2004,18 (14): 1734-1736.
28 Jordan P, Cunha C, Carmo-Fonseca M. The cdk7-cyclin H-MAT1 complex associated with TFIIH is localized in coiled bodies. Mol Biol Cell. 1997,8 (7):1-207-1217.
29 Tassan J P, Schultz S J, Bartek J, et al. Cell cycle analysis of the activity, subcellular localization, and subunit composition of human CAK (CDK-activating kinase). J Cell Biol. 1994,127 (2):467-478.
30 Lamond A 1, Earnshaw W C. Structure and function in the nucleus. Science. 1998,280 (5363):547-553.
31 Faust M, Kartarius S, Schwindling S L, et al. Cyclin H is a new binding partner for protein kinase CK2. Biochem Biophys Res Commun. 2002,296 (1): 13-19.
32 Gannon J V, Nebreda A, Goodger N M, et al. A measure of the mitotic index: studies of the abundance and half-life of p34cdc2 in cultured cells and normal and neoplastic tissues. Genes Cells. 1998,3 (l):17-27.
33 Kim J M, McGaughy J T, Bogle R K, et al. Meiotic expression of the cyclin H/Cdk7 complex in male germ cells of the mouse. Biol Reprod. 2001,64 (5): 1400-1408.
34 Kaldis P, Solomon M J. Analysis of CAK activities from human cells. Eur J Biochem. 2000,267 (13):4213-4221.
35 Leclerc V, Raisin S, Leopold P. Dominant-negative mutants reveal a role for the Cdk7 kinase at the mid-blastula transition in Drosophila embryos. Embo J. 2000,19 (7):1567-1575.
36 Hermand D, Pihlak A, Westerling T, et al. Fission yeast Cskl is a
引文
1 Fcsquet D, Labbe J C, Derancourt J, et al. The MO15 gene encodes the catalytic subunit of a protein kinase that activates cdc2 and other cyclin-dependent kinases (CDKs) through phosphorylation of Thr161 and its homologues. Embo J. 1993,12 (8):3111-3121.
2 Poon R Y, Yamashita K, Adamczewski J P, et al. The cdc2-related protein p40MO15 is the catalytic subunit of a protein kinase that can activate p33cdk2 and p34cdc2. Embo J. 1993,12 (8):3123-3132.
3 Solomon M J, Harper J W, Shuttleworth J. CAK, the p34cdc2 activating kinase, contains a protein identical or closely related to p40MO15. Embo J. 1993,12 (8):3133-3142.
4 Labbe J C, Martinez A M, Fesquet D, et al. p40MO15 associates with a p36 subunit and requires both nuclear translocation and Thr176 phosphorylation to generate cdk-activating kinase activity in Xenopus oocytes. Embo J. 1994,13 (21):5155-5164.
5 Fisher R P, Morgan D O. A novel cyclin associates with MO15/CDK7 to form the CDK-activating kinase. Cell. 1994,78 (4):713-724.
6 Tassan J P, Jaquenoud M, Fry A M, et al. In vitro assembly of a functional human CDK7-cyclin H complex requires MAT1, a novel 36 kDa RING finger protein. EmboJ. 1995,14 (22):5608-5617.
7 Shiekhattar R, Mermelstein F, Fisher R P, et al. Cdk-activating kinase complex is a component of human transcription factor TFIIH. Nature. 1995,374 (6519):283-287.
8 Kaldis P. The cdk-activating kinase (CAK): from yeast to mammals. Cell Mol Life Sci. 1999,55 (2):284-296.
9 Higashi H, Suzuki-Takahashi I, Saitoh S, et al. Cyclin-dependent kinase-2 (Cdk2)forms an inactive complex with cyclin D1 since Cdk2 associated with cyclin Dl is not phosphorylated by Cdk7-cyclin-H. Eur J Biochem. 1996,237 (2):460-467.
10 Ko L J, Shieh S Y, Chen X, et al. p53 is phosphorylated by CDK7-cyclin H in a p36MATl-dependent manner. MolCell Biol. 1997,17 (12):7220-7229.
11 Roy R, Adamczewski J P, Seroz T, et al. The MO15 cell cycle kinase is associated with the TFIIH transcription-DNA repair factor. Cell. 1994,79 (6): 1093-1101.
12 Makela T P, Parvin J D, Kim J, et al. A kinase-deficient transcription factor TFIIH is functional in basal and activated transcription. Proc Natl Acad Sci US A. 1995,92 (11):5174-5178.
13 Akoulitchev S, Makela T P, Weinberg R A, et al. Requirement for TFIIH kinase activity in transcription by RNA polymerase II. Nature. 1995,377 (6549):557-560.
14 Adamczewski J P, Rossignol M, Tassan J P, et al. MAT1, cdk7 and cyclin H form a kinase complex which is UV light-sensitive upon association with TFIIH. Embo J. 1996,15 (8): 1877-1884.
15 Fisher R P. Secrets of a double agent: CDK7 in cell-cycle control and transcription. J Cell Sci. 2005,118 (Pt.22):5171-5180.
16 Larochelle S, Batliner J, Gamble M J, et al. Dichotomous but stringent substrate selection by the dual-function Cdk7 complex revealed by chemical genetics. Nat Struct Mo! Biol. 2005,
17 Kaldis P, Russo A A, Chou H S, et al. Human and yeast cdk-activating kinases (CAKs) display distinct substrate specificities. Mol Biol Cell. 1998,9 (9):2545-2560.
18 Bour G, Gaillard E, Bruck N, et al. Cyclin H binding to the RARalpha activation function (AF)-2 domain directs phosphorylation of the AF-1 domain by cyclin-dependent kinase 7. Proc Natl Acad Sci USA. 2005,102 (46): 16608-16613.
19 Devault A, Martinez A M, Fesquet D, et al. MAT1 ('menage a trois') a new RING finger protein subunit stabilizing cyclin H-cdk7 complexes in starfish and Xenopus CAK. Embo J. 1995,14 (20):5027-5036.
20 Martinez A M, Afshar M, Martin F, et al. Dual phosphorylation of the T-loop in cdk7: its role in controlling cyclin H binding and CAK activity. Embo J. 1997,16 (2):343-354.
21 Brown A J, Jones T, Shuttleworth J. Expression and activity of p40MO15, the catalytic subunit of cdk-activating kinase, during Xenopus oogenesis and embryogenesis. Mol Biol Cell. 1994,5 (8):921-932.
22 Krempler A, Kartarius S, Gunther J, et al. Cyclin H is targeted to the nucleus by C-terminal nuclear localization sequences. Cell Mol Life Sci. 2005,62 (12):1379-1387.
23 Schneider E, Kartarius S, Schuster N, et al. The cyclin H/cdk7/Matl kinaseactivity is regulated by CK2 phosphorylation of cyclin H. Oncogene. 2002,21 (33):5031-5037.
24 Wu L, Chen P, Shum C H, et al. MAT 1-modulated CAK activity regulates cell cycle G(l) exit. Mol Cell Biol. 2001,21 (l):260-270.
25 Kato J Y, Matsuoka M, Strom D K, et al. Regulation of cyclin D-dependent kinase 4 (cdk4) by cdk4-activating kinase. Mol Cell Biol. 1994,14 (4) :2713-2721.
26 Gerber M R, Farrell A, Deshaies R J, et al. Cdc37 is required for association of the protein kinase Cdc28 with Gl andmito.tic cyclins. Proc Nat! Acad Sci USA. 1995,92 (10):4651-4655.
27 He Q, Peng H, Collins S J, et al. Retinoid-modulated MAT1 ubiquitination and CAK activity. Faseb J. 2004,18 (14): 1734-1736.
28 Jordan P, Cunha C, Carmo-Fonseca M. The cdk7-cyclin H-MAT1 complex associated with TFIIH is localized in coiled bodies. Mol Biol Cell. 1997,8 (7):1-207-1217.
29 Tassan J P, Schultz S J, Bartek J, et al. Cell cycle analysis of the activity, subcellular localization, and subunit composition of human CAK (CDK-activating kinase). J Cell Biol. 1994,127 (2):467-478.
30 Lamond A 1, Earnshaw W C. Structure and function in the nucleus. Science. 1998,280 (5363):547-553.
31 Faust M, Kartarius S, Schwindling S L, et al. Cyclin H is a new binding partner for protein kinase CK2. Biochein Biophys Res Commun. 2002,296 (1): 13-19.
32 Gannon J V, Nebreda A, Goodger N M, et al. A measure of the mitotic index: studies of the abundance and half-life of p34cdc2 in cultured cells and normal and neoplastic tissues. Genes Cells. 1998,3 (l):17-27.
33 Kim J M, McGaughy J T, Bogle R K, et al. Meiotic expression of the cyclin H/Cdk7 complex in male germ cells of the mouse. Biol Reprod. 2001,64 (5): 1400-1408.
34 Kaldis P, Solomon M J. Analysis of CAK activities from human cells. Eur J Biochem. 2000,267 (13):4213-4221.
35 Leclerc V, Raisin S, Leopold P. Dominant-negative mutants reveal a role for the Cdk7 kinase at the mid-blastula transition in Drosophila embryos. Embo J. 2000,19 (7):1567-1575.
36 Hermand D, Pihlak A, Westerling T, et al. Fission yeast Cskl is aCAK-activating kinase (CAKAK). Embo J. 1998,17 (24):7230-7238.
37 Lee K M, Saiz J E, Barton W A, et al. Cdc2 activation in fission yeast depends on Mcs6 and Cskl, two partially redundant Cdk-activating kinases (CAKs). Curr Biol. 1999,9 (8):441-444.
38 Kaldis P, Sutton A, Solomon M J. The Cdk-activating kinase (CAK) from budding yeast. Cell. 1996,86 (4):553-564.
39 Kimmelman J, Kaldis P, Hengartner C J, et al. Activating phosphorylation of the Kin28p subunit of yeast TFIIH by Caklp. Mol Cell Biol. 1999,19 (7):4774-4787.
40 Feaver W J, Svejstrup J Q, Henry N L, et al. Relationship of CDK-activating kinase and RNA polymerase II CTD kinase TFI1H/TFIIK. Cell. 1994,79 (6): 1103-1109.
41 Serizawa H, Makela T P, Conaway J W, et al. Association of Cdk-activating kinase subunits with transcription factor TFIIH. Nature. 1995,374 (6519):280-282.
42 Cismowski M J, Laff G M, Solomon M J, et al. KIN28 encodes a C-terminal domain kinase that controls mRNA transcription in Saccharomyces cerevisiae but lacks cycl in-dependent kinase-activating kinase (CAK) activity. Mol Cell Biol. 1995,15 (6):2983-2992.
43 Senderowicz A M. Small-molecule cyclin-dependent kinase modulators. Oncogene. 2003,22 (42):6609-6620.
44 Dai Y, Grant S. Cyclin-dependent kinase inhibitors. Curr Opin Pharmacol. 2003,3 (4):362-370.
45 Lolli G Lowe E D, Brown N R, et al. The crystal structure of human CDK7 and its protein recognition properties. Structure (Camb). 2004,12 (11):2067-2079.
46 Lolli G Johnson LM. CAK-Cyclin-dependent Activating Kinase: a key kinase in cell cycle control and a target for drugs? Cell Cycle. 2005,4 (4):572-577.
47 Hirai H, Kawanishi N, Iwasawa Y. Recent advances in the development of selective small molecule inhibitors for cyclin-dependent kinases. Curr Top Med Chew. 2005,5 (2): 167-179.
48 Agbottah E, de La Fuente C, Nekhai S, et al. Antiviral activity of CYC202 in HIV-l -infected cells. J Biol Chem. 2005,280 (4):3029-3042.
49 Liang Y C, Tsai S H, Chen L, et al. Resveratrol-induced G2 arrest through the inhibition of CDK7 and p34CDC2 kinases in colon carcinoma HT29 cells. Biochem Pharmacol. 2003,65 (7): 1053-1060.
50 Rossi D J, Londesborough A, Korsisaari N, et al. Inability to enter S phase and defective RNA polymerase II CTD phosphorylation in mice lacking Matl. Embo J. 2001,20 (11):2844-2856.1 J. Newport, M. Kirschner, A major developmental transition in early Xenopus embryos: Ⅰ. Characterization and timing of cellular changes at the midblastula stage, Cell 30 (1982)675-686.
2 J. Newport, M. Kirschner, A major developmental transition in early Xenopus embryos:Ⅱ. Control of the onset of transcription, Cell 30 (1982) 687-696.
3 D. A. Kane, C.B. Kimmel, The zebrafish midblastula transition, Development 119 (1993) 447-456.
4 Amsterdam, A., Burgess, S., Golling, G., Chen, W., Sun, Z., Townsend, K., Farrington, S., Haldi, M., Hopkins, N., A large-scale insertional mutagenesis screen in zebrafish. Genes Dev. 1999 13, 2713-2724.
5 Schena M, Shalon D, Davis RW et al. Quantitative monitoring ofgene expression patterns with complementary DNA microarray. Science, 1995, 270(20): 467-470.
6 贾方钧，王剑伟，吴清江。异源精子诱导稀有鮈鲫的人工雌核发育。水生生物学报，2002 (3)：78-85。
7 Ronan T. Bree, Sarah McLoughlin, Suk-Won Jin, Oonagh M. McMeel, DidierY. R. Stainier, Maura Grealy, Lucy Byrnes. nanor, a novel zygotic gene, is expressed initially at the midblastula transition in zebrafish. Biochemical and Biophysical Research Communications, 2005 333: 722-728.
8 李书鸿，郭绍东，孙方臻。通过mRNA差异显示分离一个新的斑马鱼胚胎发育基因。科学通报，1998，43 (7)：759-764。
9 Sanada, Y. Oue, N. Mitani, Y. Yoshida, K. Nakayama, H. Yasui, W., Downregulation of the claudin-18 gene, identified through serial analysis of gene expression data analysis, in gastric cancer with an intestinal phenotype. J Pathol 2006.
10 Elwood Linneya, Betsy Dobbs-McAuliffeb, Hedieh Sajadic, Renae L. Malek, Microarray gene expression' profiling during the segmentation phase of zebrafish Development. Comparative Biochemistry and Physiology, Part C2004 138: 351-362.
11 Kamme K Salunga R, Yu J, Tran DT, Zhu J, Luo L, Bittner A, Guo HQ, Miller N, Wan J, Erlander M. Single-cell microarray analysis in hippocampus CAI: demonstration and validation of cellular heterogeneity. J Neurosci 2003;??23(9): 3607～3615.
12 Dixon AK, Richardson PJ, Pinnock RD, Lee K. Gene-expression analysis at the single-cell level. Trends Pharmacol Sci 2000; 21(2): 65～70.
13 Sbih JH, Michalowska AM, Dobbin K, Ye Y, Qiu TH, Green JE. Effects of pooling mRNA in microarray class comparisons. Bioinformatics 2004; 20(18): 3318～3325.
14 Kendziorski CM, Zhang Y, Lan H, Attie AD. The efficiency of pooling mRNA in microarray experiments. Biostatistics 2003; 4(3):465～477.1 徐振华等。生命的化学，2002，22：376
2 Westerfield M. University of Oregon Press, 1993
3 Hu XY et al. Cell Res, 2002, 12157
4 朱立平等。免疫学常用实验方法，第一版，北京：人民军医出版社，2000，352
5 Gu J et al. BioTechniques, 1994, 17: 257
2 Poon R Y, Yamashita K, Adamczewski J P, et al. The cdc2-related protein p40MO15 is the catalytic subunit of a protein kinase that can activate p33cdk2 and p34cdc2. Embo J. 1993,12 (8):3123-3132.
3 Solomon M J, Harper J W, Shuttleworth J. CAK, the p34cdc2 activating kinase, contains a protein identical or closely related to p40MO15. Embo J. 1993,12 (8):3133-3142.
4 Labbe J C, Martinez A M, Fesquet D, et al. p40MO15 associates with a p36 subunit and requires both nuclear translocation and Thr176 phosphorylation to generate cdk-activating kinase activity in Xenopus oocytes. Embo J. 1994,13 (21):5155-5164.
5 Fisher R P, Morgan D O. A novel cyclin associates with MO15/CDK7 to form the CDK-activating kinase. Cell. 1994,78 (4):713-724.
6 Tassan J P, Jaquenoud M, Fry A M, et al. In vitro assembly of a functional human CDK7-cyclin H complex requires MAT1, a novel 36 kDa RING finger protein. EmboJ. 1995,14 (22):5608-5617.
7 Shiekhattar R, Mermelstein F, Fisher R P, et al. Cdk-activating kinase complex is a component of human transcription factor TFIIH. Nature. 1995,374 (6519):283-287.
8 Kaldis P. The cdk-activating kinase (CAK): from yeast to mammals. Cell Mol Life Sci. 1999,55 (2):284-296.
9 Higashi H, Suzuki-Takahashi I, Saitoh S, et al. Cyclin-dependent kinase-2 (Cdk2)forms an inactive complex with cyclin D1 since Cdk2 associated with cyclin Dl is not phosphorylated by Cdk7-cyclin-H. Eur J Biochem. 1996,237 (2):460-467.
10 Ko L J, Shieh S Y, Chen X, et al. p53 is phosphorylated by CDK7-cyclin H in a p36MATl-dependent manner. MolCell Biol. 1997,17 (12):7220-7229.
11 Roy R, Adamczewski J P, Seroz T, et al. The MO15 cell cycle kinase is associated with the TFIIH transcription-DNA repair factor. Cell. 1994,79 (6): 1093-1101.
12 Makela T P, Parvin J D, Kim J, et al. A kinase-deficient transcription factor TFIIH is functional in basal and activated transcription. Proc Natl Acad Sci US A. 1995,92 (11):5174-5178.
13 Akoulitchev S, Makela T P, Weinberg R A, et al. Requirement for TFIIH kinase activity in transcription by RNA polymerase II. Nature. 1995,377 (6549):557-560.
14 Adamczewski J P, Rossignol M, Tassan J P, et al. MAT1, cdk7 and cyclin H form a kinase complex which is UV light-sensitive upon association with TFIIH. Embo J. 1996,15 (8): 1877-1884.
15 Fisher R P. Secrets of a double agent: CDK7 in cell-cycle control and transcription. J Cell Sci. 2005,118 (Pt.22):5171-5180.
16 Larochelle S, Batliner J, Gamble M J, et al. Dichotomous but stringent substrate selection by the dual-function Cdk7 complex revealed by chemical genetics. Nat Struct Mo! Biol. 2005,
17 Kaldis P, Russo A A, Chou H S, et al. Human and yeast cdk-activating kinases (CAKs) display distinct substrate specificities. Mol Biol Cell. 1998,9 (9):2545-2560.
18 Bour G, Gaillard E, Bruck N, et al. Cyclin H binding to the RARalpha activation function (AF)-2 domain directs phosphorylation of the AF-1 domain by cyclin-dependent kinase 7. Proc Natl Acad Sci USA. 2005,102 (46): 16608-16613.
19 Devault A, Martinez A M, Fesquet D, et al. MAT1 ('menage a trois') a new RING finger protein subunit stabilizing cyclin H-cdk7 complexes in starfish and Xenopus CAK. Embo J. 1995,14 (20):5027-5036.
20 Martinez A M, Afshar M, Martin F, et al. Dual phosphorylation of the T-loop in cdk7: its role in controlling cyclin H binding and CAK activity. Embo J. 1997,16 (2):343-354.
21 Brown A J, Jones T, Shuttleworth J. Expression and activity of p40MO15, the catalytic subunit of cdk-activating kinase, during Xenopus oogenesis and embryogenesis. Mol Biol Cell. 1994,5 (8):921-932.
22 Krempler A, Kartarius S, Gunther J, et al. Cyclin H is targeted to the nucleus by C-terminal nuclear localization sequences. Cell Mol Life Sci. 2005,62 (12):1379-1387.
23 Schneider E, Kartarius S, Schuster N, et al. The cyclin H/cdk7/Matl kinaseactivity is regulated by CK2 phosphorylation of cyclin H. Oncogene. 2002,21 (33):5031-5037.
24 Wu L, Chen P, Shum C H, et al. MAT 1-modulated CAK activity regulates cell cycle G(l) exit. Mol Cell Biol. 2001,21 (l):260-270.
25 Kato J Y, Matsuoka M, Strom D K, et al. Regulation of cyclin D-dependent kinase 4 (cdk4) by cdk4-activating kinase. Mol Cell Biol. 1994,14 (4) :2713-2721.
26 Gerber M R, Farrell A, Deshaies R J, et al. Cdc37 is required for association of the protein kinase Cdc28 with Gl andmito.tic cyclins. Proc Nat! Acad Sci USA. 1995,92 (10):4651-4655.
27 He Q, Peng H, Collins S J, et al. Retinoid-modulated MAT1 ubiquitination and CAK activity. Faseb J. 2004,18 (14): 1734-1736.
28 Jordan P, Cunha C, Carmo-Fonseca M. The cdk7-cyclin H-MAT1 complex associated with TFIIH is localized in coiled bodies. Mol Biol Cell. 1997,8 (7):1-207-1217.
29 Tassan J P, Schultz S J, Bartek J, et al. Cell cycle analysis of the activity, subcellular localization, and subunit composition of human CAK (CDK-activating kinase). J Cell Biol. 1994,127 (2):467-478.
30 Lamond A 1, Earnshaw W C. Structure and function in the nucleus. Science. 1998,280 (5363):547-553.
31 Faust M, Kartarius S, Schwindling S L, et al. Cyclin H is a new binding partner for protein kinase CK2. Biochein Biophys Res Commun. 2002,296 (1): 13-19.
32 Gannon J V, Nebreda A, Goodger N M, et al. A measure of the mitotic index: studies of the abundance and half-life of p34cdc2 in cultured cells and normal and neoplastic tissues. Genes Cells. 1998,3 (l):17-27.
33 Kim J M, McGaughy J T, Bogle R K, et al. Meiotic expression of the cyclin H/Cdk7 complex in male germ cells of the mouse. Biol Reprod. 2001,64 (5): 1400-1408.
34 Kaldis P, Solomon M J. Analysis of CAK activities from human cells. Eur J Biochem. 2000,267 (13):4213-4221.
35 Leclerc V, Raisin S, Leopold P. Dominant-negative mutants reveal a role for the Cdk7 kinase at the mid-blastula transition in Drosophila embryos. Embo J. 2000,19 (7):1567-1575.
36 Hermand D, Pihlak A, Westerling T, et al. Fission yeast Cskl is aCAK-activating kinase (CAKAK). Embo J. 1998,17 (24):7230-7238.
37 Lee K M, Saiz J E, Barton W A, et al. Cdc2 activation in fission yeast depends on Mcs6 and Cskl, two partially redundant Cdk-activating kinases (CAKs). Curr Biol. 1999,9 (8):441-444.
38 Kaldis P, Sutton A, Solomon M J. The Cdk-activating kinase (CAK) from budding yeast. Cell. 1996,86 (4):553-564.
39 Kimmelman J, Kaldis P, Hengartner C J, et al. Activating phosphorylation of the Kin28p subunit of yeast TFIIH by Caklp. Mol Cell Biol. 1999,19 (7):4774-4787.
40 Feaver W J, Svejstrup J Q, Henry N L, et al. Relationship of CDK-activating kinase and RNA polymerase II CTD kinase TFI1H/TFIIK. Cell. 1994,79 (6): 1103-1109.
41 Serizawa H, Makela T P, Conaway J W, et al. Association of Cdk-activating kinase subunits with transcription factor TFIIH. Nature. 1995,374 (6519):280-282.
42 Cismowski M J, Laff G M, Solomon M J, et al. KIN28 encodes a C-terminal domain kinase that controls mRNA transcription in Saccharomyces cerevisiae but lacks cycl in-dependent kinase-activating kinase (CAK) activity. Mol Cell Biol. 1995,15 (6):2983-2992.
43 Senderowicz A M. Small-molecule cyclin-dependent kinase modulators. Oncogene. 2003,22 (42):6609-6620.
44 Dai Y, Grant S. Cyclin-dependent kinase inhibitors. Curr Opin Pharmacol. 2003,3 (4):362-370.
45 Lolli G Lowe E D, Brown N R, et al. The crystal structure of human CDK7 and its protein recognition properties. Structure (Camb). 2004,12 (11):2067-2079.
46 Lolli G Johnson LM. CAK-Cyclin-dependent Activating Kinase: a key kinase in cell cycle control and a target for drugs? Cell Cycle. 2005,4 (4):572-577.
47 Hirai H, Kawanishi N, Iwasawa Y. Recent advances in the development of selective small molecule inhibitors for cyclin-dependent kinases. Curr Top Med Chew. 2005,5 (2): 167-179.
48 Agbottah E, de La Fuente C, Nekhai S, et al. Antiviral activity of CYC202 in HIV-l -infected cells. J Biol Chem. 2005,280 (4):3029-3042.
49 Liang Y C, Tsai S H, Chen L, et al. Resveratrol-induced G2 arrest through the inhibition of CDK7 and p34CDC2 kinases in colon carcinoma HT29 cells. Biochem Pharmacol. 2003,65 (7): 1053-1060.
50 Rossi D J, Londesborough A, Korsisaari N, et al. Inability to enter S phase and defective RNA polymerase II CTD phosphorylation in mice lacking Matl. Embo J. 2001,20 (11):2844-2856.1 J. Newport, M. Kirschner, A major developmental transition in early Xenopus embryos: Ⅰ. Characterization and timing of cellular changes at the midblastula stage, Cell 30 (1982)675-686.
2 J. Newport, M. Kirschner, A major developmental transition in early Xenopus embryos:Ⅱ. Control of the onset of transcription, Cell 30 (1982) 687-696.
3 D. A. Kane, C.B. Kimmel, The zebrafish midblastula transition, Development 119 (1993) 447-456.
4 Amsterdam, A., Burgess, S., Golling, G., Chen, W., Sun, Z., Townsend, K., Farrington, S., Haldi, M., Hopkins, N., A large-scale insertional mutagenesis screen in zebrafish. Genes Dev. 1999 13, 2713-2724.
5 Schena M, Shalon D, Davis RW et al. Quantitative monitoring ofgene expression patterns with complementary DNA microarray. Science, 1995, 270(20): 467-470.
6 贾方钧，王剑伟，吴清江。异源精子诱导稀有鮈鲫的人工雌核发育。水生生物学报，2002 (3)：78-85。
7 Ronan T. Bree, Sarah McLoughlin, Suk-Won Jin, Oonagh M. McMeel, DidierY. R. Stainier, Maura Grealy, Lucy Byrnes. nanor, a novel zygotic gene, is expressed initially at the midblastula transition in zebrafish. Biochemical and Biophysical Research Communications, 2005 333: 722-728.
8 李书鸿，郭绍东，孙方臻。通过mRNA差异显示分离一个新的斑马鱼胚胎发育基因。科学通报，1998，43 (7)：759-764。
9 Sanada, Y. Oue, N. Mitani, Y. Yoshida, K. Nakayama, H. Yasui, W., Downregulation of the claudin-18 gene, identified through serial analysis of gene expression data analysis, in gastric cancer with an intestinal phenotype. J Pathol 2006.
10 Elwood Linneya, Betsy Dobbs-McAuliffeb, Hedieh Sajadic, Renae L. Malek, Microarray gene expression' profiling during the segmentation phase of zebrafish Development. Comparative Biochemistry and Physiology, Part C2004 138: 351-362.
11 Kamme K Salunga R, Yu J, Tran DT, Zhu J, Luo L, Bittner A, Guo HQ, Miller N, Wan J, Erlander M. Single-cell microarray analysis in hippocampus CAI: demonstration and validation of cellular heterogeneity. J Neurosci 2003;??23(9): 3607～3615.
12 Dixon AK, Richardson PJ, Pinnock RD, Lee K. Gene-expression analysis at the single-cell level. Trends Pharmacol Sci 2000; 21(2): 65～70.
13 Sbih JH, Michalowska AM, Dobbin K, Ye Y, Qiu TH, Green JE. Effects of pooling mRNA in microarray class comparisons. Bioinformatics 2004; 20(18): 3318～3325.
14 Kendziorski CM, Zhang Y, Lan H, Attie AD. The efficiency of pooling mRNA in microarray experiments. Biostatistics 2003; 4(3):465～477.1 徐振华等。生命的化学，2002，22：376
2 Westerfield M. University of Oregon Press, 1993
3 Hu XY et al. Cell Res, 2002, 12157
4 朱立平等。免疫学常用实验方法，第一版，北京：人民军医出版社，2000，352
5 Gu J et al. BioTechniques, 1994, 17: 257