斑节对虾GTF2H4基因的分子克隆及表达分析

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

基础转录因子IIH亚基4（generaltranscriptionfactorIIHsubunit4，GTF2H4）作为基础转录因子IIH（TFI—IH）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（Penaeusmonodor）为研究对象，利用eDNA末端快速扩增技术（rapidamplicationofeDNAends，RACE）获得了PmGTF2H4基因的cDNA全长。该序列全长1401bp，可编码466个氨基酸，其中包括108bp的5’非编码区域（UTR）和372bp的3’UTR。同源性分析显示，PmGTF2H4与阿里山潜蝇茧蜂（Fopiusarsanus）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6～48h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。

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 of GTF2H4 was cloned from black tiger shrimp （Penaeus monodon） by rapid amplification of cDNA ends （RACE） approaches. The full-length cDNA of black tiger shrimp GTF2H4 （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 reported GTF2H4 genes. QRT-PCR indicated that the temporal expression of PmGTF2H4 in the ovaries was significantly higher than that in the other tissues （P 〈 0. 05 ）. PmGTF2H4 was predominantly detectable in stage V ovaries. The injection of 5-HT （50 μg ·g-1 body weight） promoted the expression level of PmGTF2H4 in the ovaries at 6 -48 h post injection （hpi）. These results indicate that PmGTF2H4 might be involved in the regulation