籽鹅促卵泡激素α亚基基因的克隆、序列分析及其原核表达载体的构建

以雌性籽鹅脑垂体的总RNA为模板，利用特异性引物通过RT-PCR扩增获得长为363 bp的籽鹅促卵泡激素α亚基的cDNA片段，将扩增的促卵泡激素α亚基基因片段克隆至pMD18-T载体后进行测序。将测序结果与鸡、鹌鹑、火鸡、鼠、羊、牛等多种禽类和哺乳动物的该基因核苷酸序列及推导的氨基酸序列进行同源性分析。结果表明，这些物种促卵泡激素α亚基基因序列具有较高的保守性，其中与鸡、鹌鹑、火鸡的核苷酸同源性最高，均为95.9%，推导的氨基酸序列与鸡、鹌鹑、火鸡的同源性最高，均为97.5%。为了对克隆的籽鹅促卵泡激素α亚基基因功能研究提供基础，将籽鹅促卵泡激素α亚基基因克隆至pET-32a(+)原核表达载体。

Cloning,Sequence Analysis and Construction of Prokaryotic Expression Vector of the FSH Alpha Subunit Gene in Zi Goose

The total RNA was extracted from the pituitary of Zi goose, and the cDNA of the FSHα was amplified by RT-PCR. The amplified FSHα gene is 396 bp in length. The FSHα gene was cloned into the vector pMD18-T. The positive clones were verified and sequenced. The nucleotide sequence and deduced amino acid sequence were compared with homologous sequences from available species including chicken, quail, turkey, mouse, sheep, cattle, et al. Sequence alignment showed that FSHα gene in these species were highly conservative. FSHα gene had the highest homology of 95.9% with chicken, quail and turkey in nucleotide, and had the highest homology of 97.5% with chicken, quail and turkey in amino acids. Then the clones of FSHα gene were inserted into pET-32a (+) prokaryotic expression vector.