摘要
以鸭肠炎病毒(DEV)SD-01株DNA为模板,根据Gen Bank上已发表的基因序列设计一对引物,利用PCR技术扩增出US2全基因,将PCR产物连入pGM-T载体。用BamHI和EcoR V双酶切重组载体,使US2基因缺失约130bp,将含双loxP酶切位点的EGFP完整表达盒插入BamH I和EcoR V酶切位点之间,成功构建了以US2基因为重组臂的含EGFP基因的转移质粒载体pUS2-loxP-EGFP-loxP。
DEV SD-01 strain genomic DNA was extracted and a pair of primers was synthesized according to the published nueleotide sequence in GenBank. The US2 gene was amplified by polymerase chain reaction (PCR) with extracted total DNA from DEV SD-01 strain as template and was cloned into vector pGM-T. The EGFP expression cassette flanked by two loxP sites was inserted between BamH I and EcoR V sites of the US2 gene to give rise to the transferring vector pUS2-loxP-EGFP-loxP.
出处
《家禽科学》
2007年第11期14-16,共3页
Poultry Science