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 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 of ovarian development and maturation in black tiger shrimp.