摘要
为获得鸡C3d基因的cDNA,从鸡肝细胞提取总RNA,通过RT-PCR扩增出C3d基因的cDNA,脂糖凝胶电泳鉴定后直接克隆到pMD18-T载体,构建C3d-pMD18-T重组质粒,转化大肠杆菌,酶切鉴定后进行序列测定。通过琼脂糖凝胶电泳证明有1.0 kb左右的PCR扩增片段,序列测定表明为C3d基因的cDNA。通过RT-PCR法从鸡肝细胞RNA中成功地扩增到鸡C3d基因的cDNA,为进一步构建基于C3d基因内佐剂的疫苗奠定了基础。
To clone and identify cDNA of AA chickens C3d, we extracted total cell RNA from the liver tissue of AA chickens and the cDNA of C3d was amplified by reverse transcription polymerase chain reaction （RT-PCR）. The cDNA fragments were directly inserted into pMD18 -T plasmid. The recombinant C3d- pMD18 -T was identified by means of restriction endonu cleases digestion and sequence. The recombinant plas- mid contains about 1.0 kb insert fragment and the sequence of the insert was identical to the published sequence encoding LaiHang chickens C3d. So the AA chicken C3d cDNA was successfully cloned.
出处
《山东农业大学学报（自然科学版）》
CSCD
北大核心
2008年第4期549-552,共4页
Journal of Shandong Agricultural University：Natural Science Edition
基金
山东省财政支持项目(SDGP2004-54-O)
