鸡腺病毒内蒙古分离株六邻体蛋白基因片段的合成和分子克隆

Molecular Cloning of Hexon Gene Fragment from Fowl Adenovirus NeiMongol Strain

根据鸡１０型腺病毒以及人２、５、４０、４１型腺病毒、牛３型、鼠１型腺病毒六邻体蛋白基因序列，选择保守区，设计和合成一对引物，以鸡腺病毒内蒙古分离株基因组ＤＮＡ为模板，进行聚合酶链反应（ＰＣＲ）扩增得到预期大小的０．５５ｋｂＤＮＡ片段．将此ＤＮＡ片段克隆于ｐＵＣ１９的ＳｍａＩ位点，筛选重组质粒，进行限制酶切分析和ＰＣＲ检测，得到含有六邻体蛋白基因片段的重组质粒。

According to the nucleotide sequences of fowl adenovirus serotype 10, human adenovirus type 2, 5, 40 amd 41, bovine adenovirus type 3 and murine adenovirus type 1, a pair of primers were designed and synthesized. The predicted length 0.55 kb DNA fragment was amplified from DNA of fowl adenovirus NeiMongol strain using this pair primers by polymerase chain reaction (PCR). The DNA fragment was cloned into SmaI site of vector pUC19. After transformation the recombinant plasmids were selected by a small-scale isolation of plasmid and agarose gel electrophoresis. The obtained recombinant plasmids were further identified by restriction enzyme analysis and PCR. The cloning of DNA fragment from hexon gene is of great importance for further molecular biological studies and molecular diagnosis of fowl adenovirus.