宁乡猪生长激素基因的克隆及其原核表达载体的构建

Cloning of Ningxiang Porcine Growth Hormone Gene and Construction of Its Prokaryotic Expression Vector

[目的]研究了宁乡猪生长激素基因(nxGH)的克隆及其原核表达载体的构建。[方法]根据已报道的猪生长激素基因序列设计了1对特异性引物,应用RT"PCR技术从宁乡猪脑垂体中克隆了nxGH编码区序列。[结果]扩增片段全长651 bp,共编码216个氨基酸。该序列与已报道的三元和五指山猪生长激素cDNA基因核苷酸序列同源性为99.85%。将nxGH cDNA序列定向插入pET"32a原核表达载体中,成功构建重组原核表达质粒pET"GH。[结论]为下一步制备宁乡猪生长激素抗体奠定了基础。

[Objective] The research aimed to study on the clone of Ningxiang porcine growth hormone gene（nxGH） and the construction of its prokaryotic expression vector.[Method] One special primer was designed based on the reported porcine growth hormone gene order.The coding sequence of nxGH was cloned from pituitary of Ningxiang porcine by using RT-PCR.[Result] The entire open reading frame of the nxGH was 651 bp and codified 216 amino acid residues.The nucleotides identity between the nxGH and the reported porcine growth hormone gene was 99.85 %.The nxGH was inserted into prokaryotic expression vector（pET-32a） and the recombination vector pET-GH was constructed.[Conclusion] The study established the foundation for preparing Ningxiang porcine growth hormone antibody in future.