摘要
目的 构建反义骨桥蛋白 (OPN)基因重组质粒 ,以用于肝癌基因治疗的研究。方法 通过设计含有特定酶切位点的引物 ,以含有全长骨桥蛋白基因的质粒 (pBlueScript OPN)为模板 ,将PCR产物反向克隆至 pcDNA3 1(+)真核表达载体上 ,并经酶切鉴定及测序分析。 结果 重组质粒经酶切后出现 913bp长的片段 ,测序分析结果与文献报道结果完全一致。结论 反义骨桥蛋白基因表达质粒克隆成功 。
Objective To construct a plasmid containing anti-sense osteopontin(OPN) gene in order to use in gene therapy for hepatocellular carcinoma.Methods The coding sequence OPN fragment was amplified by reverse transcription-PCR from the plasmid pBlueScript-OPN containing full-length human osteopontin(kind gift from Dr.Chambers,Canada).The resultant BamH1-EcoR1 cDNA fragment was reversely ligated into the expression vector pcDNA3.1(+),and confirmed by restricting enzyme digestion and DNA sequencing.Results RT-PCR product is 913 bp specific segment. Conclusions The OPN fragment was successfully cloned to the vector pcDNA3.1(+).Analysis by enzyme digestion and DNA sequence showed results from restricting enzyme.
出处
《江苏医药》
CAS
CSCD
北大核心
2004年第7期485-486,共2页
Jiangsu Medical Journal
