摘要
以鸡甲状旁腺组织中总RNA为模板,采用RT-PCR法获得甲状旁腺素(PTH)cDNA。将该基因克隆至pCDNA3载体中,构建真核细胞表达载体pCDNA3-PTH,DNA测序证明获得了PTH基因,其序列与GenBank报道序列比较,核苷酸同源性为99.4%,在第159位处存在C→A的有意义突变。将测序正确的PTH基因在多聚乙肽亚胺(PEI)介导下转染COS-1细胞,通过间接免疫荧光试验(IFA)检测到了PTH在COS-1中的表达。PTH基因克隆和表达均获得成功,为进一步探讨该基因在改善蛋壳质量中的应用奠定了基础。
Total RNA was extracted from chicken parathyroid gland tissue, and then the parathyroid hormone （PTH） cDNA was obtained by RT-PCR. The PTH gene fragment was cloned into pCDNA3 vector, and thus the pCDNA3-PTH vector was constructed. The PTH gene encoding 119 amino acids was 360 bp in length, and the homologous comparison showed 99.4% with the sequence reported in GenBank （M36522. 1 ）, except that one nueleotide changed from C to A at 159 base site. The recombinant plasmid was transfected into COS-1 cells. At 72 hours after transfection, the expression of PTH was confirmed by indirect immunofluorescence assay （IFA） with the antiserum of PTH expressed in COS-1. This study laid a foundation for further applications of PTH in diagnosis and improving eggshell quality.
出处
《畜牧与兽医》
北大核心
2008年第10期1-4,共4页
Animal Husbandry & Veterinary Medicine
