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王桂苹, 杜合军, 冷小茜, 李创举, 曹宏. 中华鲟促性腺激素释放激素受体基因cDNA序列的克隆及表达分析[J]. 水生生物学报, 2018, 42(1): 94-98. DOI: 10.7541/2018.012
引用本文: 王桂苹, 杜合军, 冷小茜, 李创举, 曹宏. 中华鲟促性腺激素释放激素受体基因cDNA序列的克隆及表达分析[J]. 水生生物学报, 2018, 42(1): 94-98. DOI: 10.7541/2018.012
Gui-Ping WANG, He-Jun DU, Xiao-Qian LENG, Chuang-Ju LI, Hong CAO. CLONING AND EXPRESSION ANALYSIS OF GONADOTROPIN-RELEASING HORMONE RECEPTOR, GNRH-R IN CHINESE STURGEON (ACIPENSER SINENSIS)[J]. ACTA HYDROBIOLOGICA SINICA, 2018, 42(1): 94-98. DOI: 10.7541/2018.012
Citation: Gui-Ping WANG, He-Jun DU, Xiao-Qian LENG, Chuang-Ju LI, Hong CAO. CLONING AND EXPRESSION ANALYSIS OF GONADOTROPIN-RELEASING HORMONE RECEPTOR, GNRH-R IN CHINESE STURGEON (ACIPENSER SINENSIS)[J]. ACTA HYDROBIOLOGICA SINICA, 2018, 42(1): 94-98. DOI: 10.7541/2018.012

中华鲟促性腺激素释放激素受体基因cDNA序列的克隆及表达分析

CLONING AND EXPRESSION ANALYSIS OF GONADOTROPIN-RELEASING HORMONE RECEPTOR, GNRH-R IN CHINESE STURGEON (ACIPENSER SINENSIS)

  • 摘要: 为了研究中华鲟(Acipenser sinensis)促性腺激素释放激素受体(Gonadotropin-releasing hormone receptor, GnRH-R)基因在中华鲟中的组织表达特征, 为中华鲟生长发育调控研究提供基础数据, 通过构建中华鲟(Acipenser sinensis)垂体的SMART cDNA质粒文库, 采用cDNA末端快速扩增(RACE), 克隆得到了中华鲟GnRH-R基因的cDNA全长序列。该序列全长1530 bp, 有478 bp的5′非翻译区, 579 bp的开放阅读框和473 bp的3′非翻译区, 共编码192个氨基酸, 其成熟多肽含有5个N连糖基化位点。通过和已知其他鱼类的GnRH-R基因进行氨基酸序列多重比对, 发现其与真鲷(Pagrus major)的同源性最高, 为76%, 与米氏叶吻银鲛(Callorhinchus milii)的同源性最低, 为39%。采用实时荧光定量PCR (Real time PCR)方法, 检测了GnRH-R的mRNA在中华鲟心、肝、脾、肾、肠道、精巢、肌肉及脑组织中的表达状况, 发现其在精巢中大量转录, 而在其他组织中则表达微弱。以上结果表明中华鲟GnRH-R基因在性腺发育特别是精子发生过程中可能起重要作用。

     

    Abstract: The Chinese sturgeon (Acipenser sinensis), an endangered and national protected species, is an important resource for sturgeon aquaculture industry. This study cloned the full-length cDNA of the gonadotropin-releasing hormone receptor (GnRH-R) from pituitary cDNA library of a 24 years old female Chinese sturgeon using rapid amplification of cDNA ends (RACE). The nucleotide sequences of AsGnRH-R is 1530 bp in length, containing a 478 bp 5′-untranslated region (UTR), a 473 bp 3′-UTR, and a 579 bp open reading frame (ORF) with a mature peptide of 192 amino acids encompassing 5 putative N-glycosylation sites. Multiple sequence alignment and phylogenetic analysis of the deduced amino acid sequence of AsGnRH-R indicated that the most similar ortholog was red sea bream (Pagrus major) (76%), and that the least similar ortholog was Callorhinchus milii (39%). Real-time quantitative PCR analysis indicated that the AsGnRH-R was expressed in tissues including liver, spleen, kidney, testis, muscle, intestine, and brain with the highest level in the testis.

     

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