威宁绵羊GHR基因克隆及组织表达
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摘要
本试验以6月龄、1周岁和2周岁的威宁绵羊公母羊为研究对象,利用分子手段对威宁绵羊GHR基因编码区序列进行克隆,并进行生物信息学分析,此外,本试验同时采用实时荧光定量PCR的手段对GHR基因在威宁绵羊不同性别及不同生长阶段心脏、肝脏、脾脏、肺脏、肾脏及背最长肌6个组织中mRNA水平上的表达规律进行探究,结果显示:威宁绵羊GHR基因CDS区序列长度为1 905 bp,发现2个SNP位点。GHR基因在威宁绵羊各组织均有不同程度的表达,在不同性别、不同生长阶段相同组织中的表达具有一定的显著性差异。
This study was conducted on male and female Weining sheep aged 6 months, 1 year and 2 years,Then cloning GHR gene coding region sequence of Weining sheep, and bioinformatic analysis was carried out. In addition, the experiment used real-time qPCR method to analyze m RNA expression rule of GHR gene in heart,liver, spleen, lung, kidney and longissimus Dorsi of Weining sheep at different growth stages and different gender.The results showed: the sequence length of CDS region of GHR gene in Weining sheep was 1 905 bp, and 2 SNPs loci were found. The result showed that GHR gene was expressed in different degrees in all tissues of Weining sheep and there were significant differences in the expression of different gender and different growth stages in the same tissues.
This study was conducted on male and female Weining sheep aged 6 months, 1 year and 2 years,Then cloning GHR gene coding region sequence of Weining sheep, and bioinformatic analysis was carried out. In addition, the experiment used real-time qPCR method to analyze m RNA expression rule of GHR gene in heart,liver, spleen, lung, kidney and longissimus Dorsi of Weining sheep at different growth stages and different gender.The results showed: the sequence length of CDS region of GHR gene in Weining sheep was 1 905 bp, and 2 SNPs loci were found. The result showed that GHR gene was expressed in different degrees in all tissues of Weining sheep and there were significant differences in the expression of different gender and different growth stages in the same tissues.
引文
Hao L.H.,2008,Genetic polymorphism research of GH and GHRgene and correlation analysis on carcass quality traits in red steppes,Thesis for M.S.,Jilin University,Supervisor:Zhao Z.H.,pp.11-87(郝灵慧,2008,草原红牛GH和GHR基因遗传多态性研究及与胴体性状的相关分析,硕士学位论文,吉林大学,导师:赵志辉,pp.11-87)
Huang Z.G.,2006,Researches on the developmental changes of MRNAs expression levels of the genes related to growing and intramuscular fatin sheep,Dissertation for Ph.D.,Nanjing A-gricultural University,Supervisor:Xie Z.,pp.12-95(黄治国,2006,绵羊生长及肌内脂肪部分相关基因表达的发育性变化研究,博士学位论文,南京农业大学,导师:谢庄,pp.12-95)
Jia B.,Xi J.F.,Zhang S.Y.,Zhao Z.S.,Zhao R.X.,and Chen J.,2006,The developmental patterns of GH-R,IGF-1 and IGF-IR gene expression in sheep skin,Yichuan(Hereditas),28(9):1078-1082(贾斌,席继峰,张苏云,赵宗胜,赵茹茜,陈杰,2006,绵羊皮肤中GHR,IGF-1和IGF-IR基因表达的发育性变化及品种特点,遗传,28(9):1078-1082)
La Y.F.,2016,Cloning and expression of growth candidate genes and their SNPs association with growth traits in sheep,Thesis for M.S.,Gansu Agricultural University,Supervisor:Li F.D.,pp.4-61(喇永富,2016,绵羊生长候选基因克隆与组织表达及其SNPs与生长性状关联分析,硕士学位论文,甘肃农业大学,导师:李发第,pp.4-61)
Leung D.W.,Spencer S.A.,and Cachianes G.,1987,Growth hormone receptor and serum binding protein:purification,cloning and expression,Nature,330(6148):537
Liu Y.,Sun Z.Y.,Sun S.H.,Qiao D.Y.,and Chu K.K.,2013,Bioinformatics analysis of coding regions of GHR gene in cattle,Zhongguo Nainiu(China Dairy Cattle),(8):1-4(刘媛,孙志颖,孙少华,乔冬雨,楚康康,2013,牛GHR基因编码区生物信息学分析,中国奶牛,(8):1-4)
Lu L.,2014,Research on egg index,GH and GHR gene expression pattern during pigeon eggs hatchcing,Thesis for M.S.,Yangzhou University,Supervisor:Ding J.T.,pp.3-42(路璐,2014,肉鸽蛋形指数对孵化率的影响与孵化期间GH和GHR基因的表达规律,硕士学位论文,扬州大学,导师:丁家桐,pp.3-42)
Shi A.,2014,Correlation of GH,GHR genes polymorphism with growth traits in Tan sheep,Thesis for M.S.,Ningxia University,Supervisors:Liu S.D.,and Li Y.K.,pp.1-46(施安,2014,GH,GHR基因多态性与滩羊生长性状的关联分析,宁夏大学,导师:刘顺德,李颖康,pp.1-46)
Sun W.,Li D.,Ma Y.H.,Guan W.J.,Chu M.X.,Ding J.T.,Li B.C.,Zhang Y.F.,and Chen L.,2012,Developmental changesof gene expression of GHR and IGF-1 genes and their association analysis with meat quality traits in Hu sheep,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),45(22):4678-4687(孙伟,李达,马月辉,关伟军,储明星,丁家桐,李碧春,张有法,陈玲,2012,湖羊GHR和IGF-1基因表达的发育性变化及其与肉质性状的关联,中国农业科学,45(22):4678-4687)
Wang Z.,Shen X.Y.,Wang J.Z.,Pan D.X.,Hou Y.,and Liu R.Y.,2016,The polymorphism detection of GHR gene in Weining sheep,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(8):2027-2031(王振,申小云,王金洲,盘道兴,熊勇,刘若余,2016,威宁绵羊GHR基因多态性分析,基因组学与应用生物学,35(8):2027-2031)
Xiong L.,2015,Distribution and expression of growth hormone receptor(GHR)in different tissues of yak,Thesis for M.S.,Gansu Agricultural University,Supervisor:Yu S.J.,pp.4-55(熊力,2015,生长激素受体(GHR)在牦牛不同器官组织中的分布与表达,硕士学位论文,甘肃农业大学,导师:余四九,pp.4-55)
Zhang H.,2010,Polymorphisms of GH,GHR&Leptin genes and their relationship with body weight in the Gansu meat sheep new breed population,Thesis for M.S.,Gansu Agricultural University,Supervisors:Ma Y.J.,Li F.D.,and Cheng G.H.,pp.1-53(张浩,2010,甘肃肉羊新品种群GH,GHR&Leptin遗传变异及其与体重的相关分析,硕士学位论文,甘肃农业大学,导师:马友记,李发弟,陈国宏,pp.1-53)
Huang Z.G.,2006,Researches on the developmental changes of MRNAs expression levels of the genes related to growing and intramuscular fatin sheep,Dissertation for Ph.D.,Nanjing A-gricultural University,Supervisor:Xie Z.,pp.12-95(黄治国,2006,绵羊生长及肌内脂肪部分相关基因表达的发育性变化研究,博士学位论文,南京农业大学,导师:谢庄,pp.12-95)
Jia B.,Xi J.F.,Zhang S.Y.,Zhao Z.S.,Zhao R.X.,and Chen J.,2006,The developmental patterns of GH-R,IGF-1 and IGF-IR gene expression in sheep skin,Yichuan(Hereditas),28(9):1078-1082(贾斌,席继峰,张苏云,赵宗胜,赵茹茜,陈杰,2006,绵羊皮肤中GHR,IGF-1和IGF-IR基因表达的发育性变化及品种特点,遗传,28(9):1078-1082)
La Y.F.,2016,Cloning and expression of growth candidate genes and their SNPs association with growth traits in sheep,Thesis for M.S.,Gansu Agricultural University,Supervisor:Li F.D.,pp.4-61(喇永富,2016,绵羊生长候选基因克隆与组织表达及其SNPs与生长性状关联分析,硕士学位论文,甘肃农业大学,导师:李发第,pp.4-61)
Leung D.W.,Spencer S.A.,and Cachianes G.,1987,Growth hormone receptor and serum binding protein:purification,cloning and expression,Nature,330(6148):537
Liu Y.,Sun Z.Y.,Sun S.H.,Qiao D.Y.,and Chu K.K.,2013,Bioinformatics analysis of coding regions of GHR gene in cattle,Zhongguo Nainiu(China Dairy Cattle),(8):1-4(刘媛,孙志颖,孙少华,乔冬雨,楚康康,2013,牛GHR基因编码区生物信息学分析,中国奶牛,(8):1-4)
Lu L.,2014,Research on egg index,GH and GHR gene expression pattern during pigeon eggs hatchcing,Thesis for M.S.,Yangzhou University,Supervisor:Ding J.T.,pp.3-42(路璐,2014,肉鸽蛋形指数对孵化率的影响与孵化期间GH和GHR基因的表达规律,硕士学位论文,扬州大学,导师:丁家桐,pp.3-42)
Shi A.,2014,Correlation of GH,GHR genes polymorphism with growth traits in Tan sheep,Thesis for M.S.,Ningxia University,Supervisors:Liu S.D.,and Li Y.K.,pp.1-46(施安,2014,GH,GHR基因多态性与滩羊生长性状的关联分析,宁夏大学,导师:刘顺德,李颖康,pp.1-46)
Sun W.,Li D.,Ma Y.H.,Guan W.J.,Chu M.X.,Ding J.T.,Li B.C.,Zhang Y.F.,and Chen L.,2012,Developmental changesof gene expression of GHR and IGF-1 genes and their association analysis with meat quality traits in Hu sheep,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),45(22):4678-4687(孙伟,李达,马月辉,关伟军,储明星,丁家桐,李碧春,张有法,陈玲,2012,湖羊GHR和IGF-1基因表达的发育性变化及其与肉质性状的关联,中国农业科学,45(22):4678-4687)
Wang Z.,Shen X.Y.,Wang J.Z.,Pan D.X.,Hou Y.,and Liu R.Y.,2016,The polymorphism detection of GHR gene in Weining sheep,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(8):2027-2031(王振,申小云,王金洲,盘道兴,熊勇,刘若余,2016,威宁绵羊GHR基因多态性分析,基因组学与应用生物学,35(8):2027-2031)
Xiong L.,2015,Distribution and expression of growth hormone receptor(GHR)in different tissues of yak,Thesis for M.S.,Gansu Agricultural University,Supervisor:Yu S.J.,pp.4-55(熊力,2015,生长激素受体(GHR)在牦牛不同器官组织中的分布与表达,硕士学位论文,甘肃农业大学,导师:余四九,pp.4-55)
Zhang H.,2010,Polymorphisms of GH,GHR&Leptin genes and their relationship with body weight in the Gansu meat sheep new breed population,Thesis for M.S.,Gansu Agricultural University,Supervisors:Ma Y.J.,Li F.D.,and Cheng G.H.,pp.1-53(张浩,2010,甘肃肉羊新品种群GH,GHR&Leptin遗传变异及其与体重的相关分析,硕士学位论文,甘肃农业大学,导师:马友记,李发弟,陈国宏,pp.1-53)