1型鸭肝炎病毒基因组全长cDNA克隆的构建

摘要

参考1型鸭肝炎病毒基因组序列，设计7条引物，用RT—PCR方法扩增出覆盖整个病毒基因组三个忠实性片段，并按顺序组装进载体pBR322中，获得全长cDNA克隆。测序结果表明，该克隆与母本毒序列同源性达99．6％，并且5’端的T7启动子和3’端的Mlu I线性化位点均成功引入。%Seven primers were synthetized according to genome sequence of duck hepatitis virus （DHV）typel, and three fidelity were amplified by RT-PCR, and these DNA fragments covering the full genome of DHV typel fragments were inserted into pBR322 vector according to the genome sequence, then gained the full-length cDNA clone.Sequence homology is 99.6% with the clone and female parent virus, and T7 promotor and Mlu I site are respectively added to genome 5＇ end and 3＇ end.