4龄中华鲟垂体的EST分析(英文)
EXPRESSED SEQUENCE TAG ANALYSIS OF A 4 YEAR2OLD CHINESE STURGEON PITUITARY
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摘要: 表达序列标签(Expressed sequencetag，EST)是鉴定基因表达规律和发现新基因的一种有效的分子生物学手段。为了能在中华鲟(Acipenser sinensis Gray)中发现与生长和生殖内分泌调控相关的基因，我们构建了中华鲟垂体的SMARTcDNA质粒文库。垂体是调节生长和生殖内分泌的重要器官。在本研究中，通过测序筛选得到了944个EST克隆，将所得EST与GenBank数据库中的序列进行比对，结果表明，802(84.96％)个克隆可以找到同源序列，共代表461个基因，其中含132个已知功能基因;而142(15.04％)个克隆不能找到同源序列。研究发现，在所有基因中，阿黑皮素原基因(Proopiomelanocortin，POMC)是出现次数最高的基因，占总EST数的10.17％，显示出其在垂体中的重要地位。我们还发现了7个未知功能的基因并重点研究了其在心脏、肝脏、脾脏、肾脏、肌肉、精巢、卵巢和垂体等组织中的表达特异性。结果发现，4个基因:EG009334、EG009337、EG009338和EG009340为垂体特异性表达或垂体和卵巢特异性表达。对这些基因进一步的功能研究将有利于我们更好地了解中华鲟生长和生殖内分泌调控的分子机制。Abstract: Expressed sequence tag (EST) analysis is an efficient tool for gene discovery and for profiling geneexpression. In order to isolate specific functional genes involved in reproduction and endocrine regulation and to reveal their evolutionary mechanisms in Chinese sturgeon (Acipenser sinensis Gray), a chondrostean fish with a history of 140 million years, we constructed its pituitary cDNA library from a 4 year-old male1 A total of 944 random cloneswere sequenced and compared with sequences in GenBank database1 Among all the 944 EST clones, 802 (84196%) clones were identified as 461 known genes, and additional 142 (15104%) as unknown genes. Functional categorization indicated that the most abundantly expressed functional gene was the proopiomelanocortin (POMC), which accounted for almost10117% of the overall expression, indicating its important function in the pituitary. Interestingly, the expression patterns of 7 unknown geneswere analyzed in various tissues, such as heart, liver, spleen, kidney, muscle, testes, ovary and pituitary. Three different categories of expression patternswere observed from them1 Several unknown ESTs, such as EG009334, EG009337, EG009338 and EG009340, were detected to be pituitary-specific, orpituitary and ovary-specific genes. Further studies on their functions will be very useful for better understanding the mechanisms of sturgeon reproduction biology and endocrinology.
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Keywords:
- cDNA /
- Expressed sequence tag (EST) /
- Acipenser sinensis /
- Pituitary
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[1] Gong Z, Yan T, Liao J, et al. Rapid identification and isolation of zebrafish cDNA clones [J]. Gene, 1997, 201: 87-98
[2] Okubo K &Matsubara K. ComplementaryDNA sequence (EST) collections and the expression information of the human genome [J]. FEBS Letters, 1997, 403: 225-229
[3] Douglas S E, Gallant J W, Bullerwell C E, et al.W inter flounder expressed sequence tags: establishment of an EST database and Identification of novel fish genes [J] .M arineB iotechnology, 1999, 1: 458-464
[4] Ju Z, Karsi A, Kocabas A, et al. Transcriptome analysis of channel catfish (Ictalurus punctatus) : genes and expression profile from the brain [J]. Gene, 2000, 261: 373-382
[5] Quackenbush J, Liang F, Holt I, et al. The TIGR gene indices: reconstruction and representation of expressed gene sequences [J]. Nucleic Acids Research, 2000, 28: 141-145
[6] Liu Z J, Karsi A & Dunham R A. Development of polymorphic ESTmarkers suitable for genetic linkage mapping of catfish [J]. M arine B iotechnology, 1999, 1: 437-447
[7] Liu Z J, Li P, KocabasA, et al.Microsatellite-containing genes from the channel catfish brain: evidence of trinucleatide repeat expansion in the coding region of nucleotide excision repair gene RAD23B [J]. B iochem ical and B iophysical Research Communications, 2001, 289: 317-324
[8] He C, ChenL, SimmonsM, et al. Putative SNP discovery in interspecific hybrids of catfish by comparative EST analysis [J].Animal Genetics, 2003, 34: 445-448
[9] Poompuang S & Hallerman EM. Toward detection of quantitative trait loci and marker2assisted selection in fish [J]. Reviews Fish Science, 1997, 5: 253-277
[10] Liu Z J & Cordes J F. DNA marker technologies and their applications in aquaculture genetics [J]. Aquaculture, 2004, 238: 1-37
[11] Gong Z. Zebrafish expressed sequence tags and their applications [J]. M ethods Cell B iology, 1999, 60: 213-233
[12] Hirono I &Aoki T. Expressed sequence tags ofmedaka (Oryzias latipes) livermRNA [J].M olecularMarineB iology and B iotechnology, 1997, 6: 345-350
[13] Inoue S, Nam B H, Hirono I, et al. A survey of expressed genes in Japanese flounder (Paralichthys olivaceus) liver and spleen [J]. Molecular M arine B iology and B iotechnology, 1997, 6: 376-380
[14] Davey G C, Caplice N C, Martin S A, et al. A survey of genes in Atlantic Salmon (Salmo salar) as identified by expressed sequence tags [J]. Gene, 2001, 263: 121-130
[15] Martin SA, Caplice N C, Davey G C, et al. EST2based identification of genes expressed in the liver of adult Atlantic salmon (Salmo salar) [J]. B iochem ical and B iophysical Research Communications, 2002, 293: 578-585
[16] KarsiA, Li P, Dunham R A, et al. Transcriptional activities in the pituitaries of channel catfish before and after induced ovulation by injection of carp pituitary extract as revealed by expressed sequence tag analysis [J]. Journal of M olecular Endocrinology, 1998, 21: 121-129
[17] KarsiA, Cao D, Li P, et al. Transcriptome analysis of channel catfish (Ictalurus punctatus) : initial analysis of gene expression and microsatellite2containing cDNAs in the skin [J]. Gene, 2002, 285: 157-168
[18] Kim S, KarsiA, Dunham R A, et al. The skeletalmuscle alphaactin gene of channel catfish ( Ictalurus punctatus) and its association with piscine specific SINE elements [J]. Gene, 2000, 252: 173-181
[19] Kocabas M A, Li P, Cao D, et al. Expression profile of the channel catfish spleen: analysis of genes involved in immune functions [J]. M arine B iotechnology, 2002, 4: 526-536
[20] Chen SL, XuM Y, Hu SN, et al.Analysisof immunerelevant genes expressed in red sea bream (Chrysophrys major) spleen [J]. Aquaculture, 2004, 240: 115-130
[21] Shiue YL, WangL H, Chao T Y, et al. EST2based identification of genes expressed in the hypothalamus of adult tilapia, Oreochrom ismossambicus [J]. B iochem ical and B iophysical Research Communications, 2004, 316: 523-527
[22] Kono K, Ponpornpisit A & SakaiM. The analysis of expressed genes in head kidney of common carp Cyprinus carpioL. stimulated with peptidoglycan [J]. Aquaculture, 2004, 235: 37-52
[23] Sarropoulou E, Power D M, Magoulas A, et al. Comparative analysis and characterization of expressed sequence tags in gilthead sea bream (Sparus aurata) liver and embryos [J]. Aquaculture, 2005, 243: 69-81
[24] Zhou L, Yao B, XiaW, et al. EST2based identification of genes expressed in the hypothalamus of male orange-spotted grouper (Epinephelus coioides) [J]. Aquaculture, 2006, 256: 129-139
[25] Zhuang P &Boyd K. Ontogenetic behavior and migration of Chinese sturgeon, Acipenser sinensis [J]. Environmental B iology of Fishes, 2002, 65: 83-97
[26] Ke F E, Hu D G & Zhang GL. Observations on the gonadal regression of the spawning population of Chinese sturgeon below Gezhouba Dam [J]. Freshwater Fisheries, 1985, 15: 38-41 (in Chinese)
[27] Yi J F, Liu D H & TangD F. Preliminary report on gonad maturation process and artificialpropagation of the Chinese sturgeon in captivity [J]. Acta Hydrobiologica Sinica, 1999, 23: 85-86 (in Chinese)
[28] Chen X H, WeiQ W, YangD G, et al. Histological studies on gonadal origin and differentiation of cultured Acipenser sinensis [J]. Journal of Fisheries of China, 2004, 28: 633-639 (in Chinese)
[29] Nakanishi S, Inoue A, Kita T, et al. Nucleotide sequence of cloned cDNA for bovine corticotropin2lipotropin precursor [J]. Nature, 1979, 278: 423-427
[30] Metz J R, Peters J J & Flik G.Molecular biology and physiology of the melanocortin system in fish: A review [J]. General and Comparative Endocrinology, 2006, 148: 150-162
[31] McMillen IC, Mercer J E & Thorburn GD. Pro-opiomelanocortin mRNA levels fall in the fetal sheep pituitary before birth [J]. Journal of M olecular Endocrinology, 1988, 1: 141-145
[32] Yang K, Challis J R, Han V K, et al1 Pro2opiomelanocortin messenger RNA levels increase in the fetal sheep pituitary during late gestation [J]. Journal of Endocrinology, 1991, 131: 483-489
[33] Myers D A, Myers TR, Grober M S, et al.Levelsof corticotroph-in-releasing hormone messenger ribonucleic acid (mRNA) in the hypothalamic paraventricular nucleus and proopiomelanocortin mRNA in the anterior pituitary during late gestation in fetal sheep [J]. Endocrinology, 1993, 132: 2109-2116
[34] Matthews S G, Han X, Lu F, et al. Developmental changes in the distribution of pro2opiomelanocortin and prolactin mRNA in the pituitary of the ovine fetus and lamb [J]. Journal of M olecular Endocrinology, 1994, 13: 175-185
[35] Matthews S G & Challis J R1 Levels of pro2opiomelanocortin and prolactin mRNA in the fetal sheep pituitary following hypoxaemia and glucocorticoid treatment in late gestation [J]. Journal of Endocrinology, 1995, 147: 139-146
[36] FabbriA, Jannini E A, GnessiL, et al. Neuroendocrine control ofmale reproductive function: the opioid system as a model of control at multiple sites [J]. Journal of Steroid B iochem istry, 1989, 32: 145-150
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