山麻鸭SHH基因克隆和组织表达
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摘要
本研究旨在克隆鸭SHH基因了解其组织表达情况,以山麻鸭为试验材料,采集下丘脑、垂体和卵巢组织样品,提取RNA逆转录后进行SHH基因克隆,并采用实时荧光定量PCR方法检测其表达水平.本试验克隆获得山麻鸭SHH基因cDNA序列878 bp,包含了5′UTR 113 bp和编码序列765 bp,编码255个氨基酸.氨基酸序列同源性比对发现,山麻鸭SHH基因与其他禽类的同源性较高,在物种间的保守性高.山麻鸭SHH基因在下丘脑表达量显著高于垂体和各级卵泡组织(P<0.01);而在大黄卵泡中表达极显著低于大白卵泡和小黄卵泡(P<0.01).本研究克隆获得SHH基因部分序列并检测其组织表达规律,为后续功能研究打下基础.
The aim of this study was to clone duck SHH gene and detect its tissue expression. The hypothalamus, pituitary and ovary tissue samples were collected from Shanma duck, and RNA was extracted for reverse transcription.The SHH gene was cloning and the expression level was detected by real-time fluorescence quantitative PCR. In this study, 878 bp cDNA sequence of Shanma duck SHH gene was cloned, including 113 bp 5′ UTR and 765 bp coding sequence, encoding 255 amino acids. Amino acid sequence homology analysis showed that Shanma duck SHH gene shared high homology with other birds SHH and was highly conserved among species. The expression of Shanma duck SHH gene in hypothalamus was significantly higher than that in pituitary and follicular tissues(P<0.01), and the expression in larger yellow follicles was significantly lower than that in large white follicles and small yellow follicles(P<0.01). This study laid a foundation for subsequent functional analysis of the SHH gene.
The aim of this study was to clone duck SHH gene and detect its tissue expression. The hypothalamus, pituitary and ovary tissue samples were collected from Shanma duck, and RNA was extracted for reverse transcription.The SHH gene was cloning and the expression level was detected by real-time fluorescence quantitative PCR. In this study, 878 bp cDNA sequence of Shanma duck SHH gene was cloned, including 113 bp 5′ UTR and 765 bp coding sequence, encoding 255 amino acids. Amino acid sequence homology analysis showed that Shanma duck SHH gene shared high homology with other birds SHH and was highly conserved among species. The expression of Shanma duck SHH gene in hypothalamus was significantly higher than that in pituitary and follicular tissues(P<0.01), and the expression in larger yellow follicles was significantly lower than that in large white follicles and small yellow follicles(P<0.01). This study laid a foundation for subsequent functional analysis of the SHH gene.
引文
[1] VARJOSALO M,TAIPALE J.Hedgehog:functions and mechanisms[J].Genes & Development,2008,22(18):2454-2472.
[2] COHEN M M.The hedgehog signaling network[J].American Journal of Medical Genetics,2003,123A(1):5-28.
[3] INGHAM P W,MCMAHON A P.Hedgehog signaling in animal development:paradigms and principles[J].Genes & Development,2001,15(23):3059-3087.
[4] VARJOSALO M,TAIPALE J.Hedgehog signaling[J].Journal of Cell Science,2007,120(1):3-6.
[5] ECHELARD Y,EPSTEIN DJ,ST-JACQUES B,et al.Sonic hedgehog,a member of a family of putative signaling molecules,is implicated in the regulation of cns polarity[J].Cell,1993,75:1417-1430.
[6] JOHNSON R L,LAUFER E,RIDDLE R D,et al.Ectopic expression of Sonic hedgehog alters dorsal-ventral patterning of somites[J].Cell,1994,79(7):1165-1173.
[7] JEONG J,MAO J H,TENZEN T,et al.Hedgehog signaling in the neural crest cells regulates the patterning and growth of facial primordia[J].Genes & Development,2004,18(8):937-951.
[8] MIURA H,KUSAKABE Y,SUGIYAMA C,et al.Shh and Ptc are associated with taste bud maintenance in the adult mouse[J].Mechanism Development,2001,106(1-2):143-145.
[9] GODDEERIS M M,RHO S,PETIET A,et al.Intracardiac septation requires hedgehog-dependent cellular contributions from outside the heart[J].Development,2008,135(10):1887-1895.
[10] BERMAN D M,DESAI N,WANG X,et al.Roles for hedgehog signaling in androgen production and prostate ductal morphogenesis[J].Development Biology,2004,267(2):387-398.
[11] POWNALL M E.More to patterning than sonic hedgehog[J].Bioessay,1994,16(6):381-383.
[12] BüRGLIN T R.The Hedgehog protein family[J].Genome Biology,2008,9(11):241.
[13] 张昌盛,刘群,李湘涛,等.多鳍鱼Shh基因克隆与同源性分析[J].科技导报,2007(12):23-27.
[14] MARTí E,TAKADA R,BUMCROT D A,et al.Distribution of sonic hedgehog peptides in the developing chick and mouse embryo[J].Development,1995,121(8):2537-2547.
[15] 杨永彬,刘玲,王炎秋,等.卵巢和卵巢肿瘤中Hedgehog信号通路的表达[J].上海交通大学学报(医学版),2009,29(4):426-430.
[16] 孙艳.2,5-己二酮致人卵巢颗粒细胞凋亡及Sonic hedgehog信号通路调控机制的研究[D].福州:福建医科大学,2012.
[2] COHEN M M.The hedgehog signaling network[J].American Journal of Medical Genetics,2003,123A(1):5-28.
[3] INGHAM P W,MCMAHON A P.Hedgehog signaling in animal development:paradigms and principles[J].Genes & Development,2001,15(23):3059-3087.
[4] VARJOSALO M,TAIPALE J.Hedgehog signaling[J].Journal of Cell Science,2007,120(1):3-6.
[5] ECHELARD Y,EPSTEIN DJ,ST-JACQUES B,et al.Sonic hedgehog,a member of a family of putative signaling molecules,is implicated in the regulation of cns polarity[J].Cell,1993,75:1417-1430.
[6] JOHNSON R L,LAUFER E,RIDDLE R D,et al.Ectopic expression of Sonic hedgehog alters dorsal-ventral patterning of somites[J].Cell,1994,79(7):1165-1173.
[7] JEONG J,MAO J H,TENZEN T,et al.Hedgehog signaling in the neural crest cells regulates the patterning and growth of facial primordia[J].Genes & Development,2004,18(8):937-951.
[8] MIURA H,KUSAKABE Y,SUGIYAMA C,et al.Shh and Ptc are associated with taste bud maintenance in the adult mouse[J].Mechanism Development,2001,106(1-2):143-145.
[9] GODDEERIS M M,RHO S,PETIET A,et al.Intracardiac septation requires hedgehog-dependent cellular contributions from outside the heart[J].Development,2008,135(10):1887-1895.
[10] BERMAN D M,DESAI N,WANG X,et al.Roles for hedgehog signaling in androgen production and prostate ductal morphogenesis[J].Development Biology,2004,267(2):387-398.
[11] POWNALL M E.More to patterning than sonic hedgehog[J].Bioessay,1994,16(6):381-383.
[12] BüRGLIN T R.The Hedgehog protein family[J].Genome Biology,2008,9(11):241.
[13] 张昌盛,刘群,李湘涛,等.多鳍鱼Shh基因克隆与同源性分析[J].科技导报,2007(12):23-27.
[14] MARTí E,TAKADA R,BUMCROT D A,et al.Distribution of sonic hedgehog peptides in the developing chick and mouse embryo[J].Development,1995,121(8):2537-2547.
[15] 杨永彬,刘玲,王炎秋,等.卵巢和卵巢肿瘤中Hedgehog信号通路的表达[J].上海交通大学学报(医学版),2009,29(4):426-430.
[16] 孙艳.2,5-己二酮致人卵巢颗粒细胞凋亡及Sonic hedgehog信号通路调控机制的研究[D].福州:福建医科大学,2012.