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李文华, 汪洋, 李志, 桂建芳, 周莉. 银鲫转录调控因子lbh-b cDNA克隆与表达分析[J]. 水生生物学报, 2018, 42(4): 673-680. DOI: 10.7541/2018.083
引用本文: 李文华, 汪洋, 李志, 桂建芳, 周莉. 银鲫转录调控因子lbh-b cDNA克隆与表达分析[J]. 水生生物学报, 2018, 42(4): 673-680. DOI: 10.7541/2018.083
LI Wen-Hua, WANG Yang, LI Zhi, GUI Jian-Fang, ZHOU Li. CLONE AND EXPRESSION ANALYSIS OF TRANSCRIPTION COFACTOR LBH-B in GIBEL CARP[J]. ACTA HYDROBIOLOGICA SINICA, 2018, 42(4): 673-680. DOI: 10.7541/2018.083
Citation: LI Wen-Hua, WANG Yang, LI Zhi, GUI Jian-Fang, ZHOU Li. CLONE AND EXPRESSION ANALYSIS OF TRANSCRIPTION COFACTOR LBH-B in GIBEL CARP[J]. ACTA HYDROBIOLOGICA SINICA, 2018, 42(4): 673-680. DOI: 10.7541/2018.083

银鲫转录调控因子lbh-b cDNA克隆与表达分析

CLONE AND EXPRESSION ANALYSIS OF TRANSCRIPTION COFACTOR LBH-B in GIBEL CARP

  • 摘要: Lbh (Limb-bud and heart)基因是脊椎动物中高度保守的转录调控因子, 在早期胚胎发育及某些人类疾病的发病过程中发挥着重要作用。我们前期在银鲫(Carassius gibelio)垂体转录组中筛选到一个在垂体中大量表达的基因lbh-b。为了进一步研究lbh基因在银鲫的表达特征, 首先采用RACE方法克隆了银鲫lbh基因家族的成员lbh-b基因(Cglbh-b)。Cglbh-b的cDNA全长1526 bp, 开放阅读框549 bp, 共编码182个氨基酸。生物信息学分析表明CgLbh-b蛋白与其他脊椎动物的Lbh蛋白同源性在68%以上, 可能也是无序蛋白质家族的成员之一。成体组织RT-PCR分析表明Cglbh-b仅在银鲫的垂体、端脑、卵巢及眼睛中表达。不同胚胎发育时期的表达分析表明, 在受精卵至原肠胚中Cglbh-b转录产物是以母源形式存在的mRNA, 其合子转录起始于尾芽期。胚胎整体原位杂交结果显示从受精后2d到受精后3d, Cglbh-b大量表达于脑和眼睛。此外, 随着卵子成熟Cglbh-b在银鲫垂体中的表达上调。这些结果暗示, Cglbh-b可能在调控银鲫脑和眼睛的发育以及卵子成熟过程中发挥着重要作用。

     

    Abstract: LBH (limb-bud and heart) is a novel high-conserved transcription cofactor in vertebrates involved in embryonic development and pathogenesis of some human disease. We screened the lbh-b gene that was abundantly expressed in the pituitary of gibel carp, and cloned the lbh-b (Cglbh-b) cDNA sequence of lbh gene family from gibel carp cDNA library by RACE-PCR. The full-length of Cglbh-b cDNA was 1526 bp with a 549 bp long open reading frame (ORF) coding a 182 amino acid protein. Bioinformatics analysis showed that CgLbh-b protein shared high homology (>68%) with other vertebrate LBH and is one of intrinsic disordered proteins.Cglbh-b was abundantly expressed in pituitary, telencephalon, ovary and eye of adult gibel carp. The expression of Cglbh-b in female pituitaries was 4.66 times higher than that in male pituitaries, and Cglbh-b in pituitary was up-regulated during the process of oocyte maturation. During the early embryonic development, maternal mRNA of Cglbh-b was detected in embryos from 4-cells stage to gastrula stage, and its transcripts were synthesized at bud stage. The results of whole mount in situ hybridization showed that Cglbh-b was distributed on the brain and eyes of embryos from 2 days post fertilization (dpf) to 3dpf. These results suggest that Cglbh-b may play important roles in brain and retina development, and reproductive regulation in oocyte maturation of gibel carp.

     

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