王艳, 傅明骏, 赵超, 周发林, 杨其彬, 邱丽华. 斑节对虾GTF2H4基因的分子克隆及表达分析[J]. 南方水产科学, 2016, 12(2): 36-43. DOI: 10.3969/j.issn.2095-0780.2016.02.006
引用本文: 王艳, 傅明骏, 赵超, 周发林, 杨其彬, 邱丽华. 斑节对虾GTF2H4基因的分子克隆及表达分析[J]. 南方水产科学, 2016, 12(2): 36-43. DOI: 10.3969/j.issn.2095-0780.2016.02.006
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WANG Yan, FU Mingjun, ZHAO Chao, ZHOU Falin, YANG Qibin, QIU Lihua. Molecular cloning and expression analysis of the GTF2H4 gene from black tiger shrimp Penaeus monodon[J]. South China Fisheries Science, 2016, 12(2): 36-43. DOI: 10.3969/j.issn.2095-0780.2016.02.006
Citation: WANG Yan, FU Mingjun, ZHAO Chao, ZHOU Falin, YANG Qibin, QIU Lihua. Molecular cloning and expression analysis of the GTF2H4 gene from black tiger shrimp Penaeus monodon[J]. South China Fisheries Science, 2016, 12(2): 36-43. DOI: 10.3969/j.issn.2095-0780.2016.02.006
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斑节对虾GTF2H4基因的分子克隆及表达分析

Molecular cloning and expression analysis of the GTF2H4 gene from black tiger shrimp Penaeus monodon

    摘要
  • 摘要: 基础转录因子IIH亚基4( general transcription factor IIH subunit 4, GTF2H4) 作为基础转录因子IIH( TFIIH) 的重要组成成员对真核生物转录过程中所涉及的细胞生长、发育、代谢等具有重要调控作用。该研究以斑节对虾( Penaeus monodon) 为研究对象, 利用cDNA 末端快速扩增技术( rapid amplication of cDNA ends, RACE) 获得了PmGTF2H4 基因的cDNA 全长。该序列全长1 401 bp, 可编码466 个氨基酸, 其中包括108 bp 的5非编码区域( UTR) 和372 bp 的3UTR。同源性分析显示,PmGTF2H4 与阿里山潜蝇茧蜂( Fopius arisanus) 等物种的 GTF2H4 具有较高的同源性。荧光定量PCR( qRT-PCR) 结果表明, PmGTF2H4 在斑节对虾卵巢中的表达量显著高于其他组织; 在卵巢的5 个不同发育时期中,PmGTF2H4 在Ⅴ期的表达量最高, 在Ⅰ 期的表达量最低。注射五羟色胺( 5-HT) 后发现PmGTF2H4的表达水平在6~48 h 显著上调。通过上述研究结果可以预测PmGTF2H4基因参与了斑节对虾卵巢的发育, 并起了一定的作用。

     

    Abstract: General transcription factor IIH subunit 4 (GTF2H4) functions as an important factor for general transcription factor IIH (TFIIH) in regulation of the cell cycle, transcription, and DNA repair. In the present study, the cDNA ofGTF2H4 was cloned fromblack tiger shrimp (Penaeus monodon) by rapid amplification of cDNA ends (RACE) approaches. The full-length cDNA of black tiger shrimpGTF2H4 (PmGTF2H4) was characterized. It was 1 881 bp in length (ORF of 1 401 bp corresponding to 466 amino acids) . BLAST X analysis indicated that the cloned gene shared high homology with other reportedGTF2H4 genes. QRT-PCR indicated that the temporal expression ofPmGTF2H4 in the ovaries was significantly higher than that in the other tissues (P 0.05) .PmGTF2H4 waspredominantly detectable in stage Ⅴ ovaries. The injection of 5-HT ( 50 gg-1 body weight) promoted the expression level ofPmGTF2H4 in the ovaries at 6～48 h post injection (hpi) . These results indicate that PmGTF2H4 might be involved in the regulation of ovarian development and maturation in black tiger shrimp.

     

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