Molecular cloning and functional analysis of the goose FSHβ gene.

The objective of this investigation was to clone goose FSH beta-subunit cDNA and to construct a FSH fusion gene to identify the function of FSH beta mRNA during stages of the breeding cycle. The FSH beta gene was obtained by reverse transcription-PCR, and the full-length FSH beta mRNA sequence was amplified by rapid-amplification of cDNA ends. FSH beta mRNA expression was detected in reproductive tissues at different stages (pre-laying, laying period, and broody period). Additionally, the expression of 4 genes known to be involved in reproduction (FSH beta, GnRH, GH, and BMP) were evaluated in COS-7 cells expressing the fusion gene (pVITRO2-FSH alpha beta-CTP). The results show that the FSH beta gene consists of a 16 base pair (bp) 5 '-untranslated region (UTR), 396bp open reading frame, and alternative 3 '-UTRs at 518bp and 780bp, respectively. qPCR analyses revealed that FSH beta mRNA is highly transcribed in reproductive tissues, including the pituitary, hypothalamus, ovaries, and oviduct. FSH beta mRNA expression increased and subsequently decreased in the pituitary, ovaries, and oviduct during the reproductive stages. Stable FSH expression was confirmed using enzyme-linked immunosorbent assays after transfection with the pVITRO2-FSH alpha beta-CTP plasmid. FSH beta, GnRH, and BMP expression increased significantly 36h and 48h after transfection with the fusion gene in COS-7 cells. The results demonstrate that the FSH beta subunit functions in the goose reproductive cycle and provides a theoretical basis for future breeding work.