黄鳝cGnRH-Ⅱ的cDNA克隆与序列分析
CLONING AND SEQUENCE ANALYSIS OF cDNA ENCODING cGnRH-II FROM RICE FIELD EEL(MONOPTERUS ALBUS)
-
摘要: 促性腺激素释放激素(Gonadotropin-releasing hormone,GnRH)是一个保守的十肽神经家族激素,在脊椎动物的性腺发育和繁殖功能的维持方面起着重要的调控作用。本文通过运用RACE和RT-PCR方法,从黄鳝脑组织中克隆得到cGnRH-ⅡcDNA全序列,其核苷酸序列长度为617bp。该cDNA编码的cGnRH-Ⅱ的前体氨基酸序列结构组成与其他物种的cGnRH-Ⅱ前体结构一致,其推导的蛋白前体长度为83个氨基酸,包括一个信号肽、cGnRH-Ⅱ十肽和一个由蛋白水解位点(Gly-Lys-Arg)连接的GnRH联接肽,其中信号肽和联接肽的长度分别为21和49个氨基酸。cGnRH-Ⅱ的氨基酸序列和其他相关物种cGnRH-Ⅱ氨基酸序列比较结果显示,cGnRH-ⅡcDNA的蛋白编码区高度保守,而非编码区的保守性程度很低。Abstract: Gonadotropin-releasing hormone (GnRH) is a conserved nerval decapeptidethat regulates the development and maintenance of reproductive function in vertebrate1The cDNA encoding the [His5Trp7Tyr8] gonadotropin-releasing hormone ( chicken-?type GnRH, cGnRH-II) precursor from the brain of rice field eel ( Monopterus albus) has been isolated and sequenced by using reverse transcription and rapid amplification of cDNA ends ( RACE) 1From the subcloning and sequence analysis of 3c-RACE and 5c-RACE product, the complete sequence of the cDNA encoding its cGnRH-II peptide was identified and determined to be 617 bp in length1The structure of the encoded peptide is identical to all other cGnRH-II reported to date. The deduced peptide is composed of a signal peptide (SP), a GnRH decapeptide and a GnRH-associated peptide ( GAP) which is connected to GnRH by a Gly-Lys-Arg sequence1The cGnRH-II cDNA encoded SP of 21 amino acids and GAP of 49 amino acids. The amino acid sequence of cGnRH-II in combining with the Gly-Lys-Arg sequence is highly conserved during evolution when compared with the corresponding regions of other GnRH precursors. However, the untranslated region of GnRH are markedly divergent in nucleotide sequences
-
-
[1] Amano M, Urano A, Aida K. Distribution and function of go-nadotropin-releasing hormone(GnRH) in the teleost brain [J]. ZoolSci, 1997, 14:1) 14
[1] [J]. Endocrinology, 2003, 144(8): 3297) 3300
[2] Matsuo H, Baba Y, Nair R M G, et al. Structure of porcine LH- andFSH-releasing hormone. I. The proposed amino acid sequence [J].Biochem Biophys Res Commun, 1971, 43: 1334) 1339
[2] [J]. Mol Cell Endocrinol, 1999, 157(1) 2): 41) 54
[3] Burgus R, BucherM, Amoss N. Primary structure of ovine hypotha-lamic luteinizing hormone-releasing factor (LRF) [J]. Proc NatlAcad Sci USA, 1972, 69: 278) 282
[4] Yamamoto N. Three gonadotropin-releasing hormone neuronal groupswith special reference to teleosts [J]. Anat Sci Internal, 2003, 78:139) 155
[5] Fernald R, White R B. Gonadotropin-releasing hormone genes: phy-logeny, structure and functions [J]. Front. Neuro., 1999, 20:224) 240
[6] Somoza G M, Miranda L A, Strob- l Mazzulla P. Gonadotropin-releas-ing hormone (GnRH): from fish to mammalian brains [J]. CellMol. Neurobiol., 2002, 22(5/6): 589) 609
[7] Lin H R. The structural variants and functional diversity of go-nadotropin-releasing hormone in vertebrates [J]. Acta ZoologicalSinica, 1998, 44(2): 226) 234 [林浩然. 脊椎动物促性腺激素释放激素的分子结构变异型和功能多样性. 动物学报,1998, 44(2): 226) 234]
[8] White R B, Eisen H A, Kasten T L, et al. Second gene for Go-nadotropin-releasing hormone in humans [J]. Proc Natl Acad SciUSA, 1998, 95: 305) 309
[9] Zhou R J, Cheng HH, Zhang Q Y, et al. SRY-related genes in thegenome of the rice field eel (Monopterus albus) [J]. Genet. Sel.Evol, 2002, 34: 129) 137
[10] Liu L, Guo Y Q, Zhou R J. The cloning and verification of the Sox 9gene of the rice field eel [J]. Acta Genetica Sinica, 2001, 28(6):535) 539 [刘利, 郭一清, 周荣家. 黄鳝 Sox9 基因的克隆及其鉴定分析. 遗传学报, 2001, 28(6): 535) 539]
[11] Tao Y X, Lin H R. Effects of exogenous hormones on serum steroidhormone in female ricefield eel (Monopterus albus) [J]. Acta Zoo-logical Sinica,1993, 39(9): 315) 321 [陶亚雄, 林浩然. 外源激素对雄性黄鳝血清类固醇激素的影响. 动物学报, 1993, 39(9): 315) 321]
[12] Gonzalez-Martinez D, Madigou T, Zmora N, et al. Differential ex-pression of three different prepro-GnRH (gonadotropin-releasing hor-mone) messengers in the brain of the European sea bass (Dicentrar-chus labrax) [J]. J Com p Neurol, 2001, 429(1): 144) 155
[13] Chow M M, Kight K E, Gothilf Y, et al. Multiple GnRHs present ina teleost species are encoded by separate genes: analysis of thesbGnRH and cGnRH- ò genes from the striped bass, Morone sax-atilis[J]. J Mol Endocrinol, 1998, 21(3): 277) 289
[14] Parhar I S, Ogawa S, Hamada T, et al. Single-cell rea- l time quant- i2期方 平等: 黄鳝 cGnRH- ò的 cDNA 克隆与序列分析263 tative polymerase chain reaction of immunofluorescently identifiedneurons of gonadotropin- releasing hormone subtypes in cichlid fish
[15] Holland M C, Gothilf Y, Meiri I, et al. Levels of the native forms ofGnRH in the pituitary of the gilthead seabream, Sparus aurata, atseveral characteristic stages of the gonadal cycle [J]. Gen Comp En-docrinol,1998, 112(3): 394) 405
[16] Okubo K, Suetake H, Aida K. A splicing variant for the prepro-mammalian gonadotropin-releasing hormone (prepro-mGnRH) mRNAis present in the brain and various peripheral tissues of the Japaneseeel [J]. Zool Sci, 1999, 16: 645) 651
[17] Okubo K, Aida K. Gonadotropin-releasing hormone gene productsdownregulate the expression of their neighboring genes that encodeprotein tyrosine phosphatases alpha and epsilon [J]. Biochem Bio-phys Res Commun, 2003, 312: 531) 536
[18] Von S K R, Harrower W L, Sherwood N M. Regulation and expres-sion of gonadotropin-releasing hormone in salmon embryo and gonad
[19] Teven C, Lehnen N, Kight K, et al. Molecular characterization ofthe GnRH system in zebrafish (Danio rerio):cloning of chickenGnRH- ò, adult brain expression patterns and pituitary content ofsalmon GnRH and chicken GnRH- ò [J]. Gen Comp Endocrinol,2003, 133(1): 27) 37
[20] Penlington M C, Williams M A, Sumpter J P, et al. Isolation andcharacterisation of mRNA encoding the salmon- and chicken- ò typegonadotropin-releasing hormones in the teleost fish Rutilus rutilus(Cyprinidae) [J]. J Mol Endocrinol, 1997, 19(3): 337) 346
[21] Ikemoto T, Park M K. Identification and characterization of the rep-tilian GnRH- ò gene in the leopard gecko, Eublepharis macularius,and its evolutionary considerations [J]. Gene, 2003, 316: 157) 165
[22] Wang L, Yoo M S, Kang H M, et al. Cloning and characterizationof cDNAs encoding the GnRH1 and GnRH2 precursors from bullfrog(Rana catesbeiana) [J]. J Exp Zool, 2001, 289(3): 190) 201
[23] Limonta P, Moretti R M, Marelli M M, et al. The biology of go-nadotropin hormone-releasing hormone: role in the control of tumorgrowth and progression in humans [J].Front Neuroendocrinol,2003, 24(4): 279) 295
[24] Gothilf Y, Elizur A, Chow M, et al. Molecular cloning and charac-terization of a novel onadotropin-releasing hormone from the giltheadseabream (Sparus aurata) [J]. Mol Mar Biol Biotechnol, 1995, 4(1):27) 35
[25] Dubois E A, Zandbergen M A, Peute H, et al. Evolutionary deve- lopment of three gonadotropin-releasing hormone (GnRH) systems invertebrates [J]. Brain Res Bull, 2002, 57: 413) 418
[26] Lin X W, Otto Carla J, Peter R E. Evolution of neuroendocrine pep-tide systems: Gonadotropin hormone-releasing hormone and somato-statin [J]. Comp. Biochem. Physiol. C, 1998, 119: 375) 388
计量
- 文章访问数: 852
- HTML全文浏览量: 0
- PDF下载量: 513
下载: