靶向骨桥蛋白shRNA表达载体的构建与鉴定

目的通过构建骨桥蛋白（osteopontin,OPN）shRNA表达载体,探索卵巢癌基因治疗新途径。方法根据基因库上的OPN—mRNA序列,设计并合成两端含有酶切位点的64个碱基的寡核苷酸链。寡核苷酸链退火后用T4DNA连接酶连接到线性化的pSilence质粒中,并对重组质粒进行酶切及序列鉴定。结果OPNsiRNA表达载体克隆构建成功,插入片段测序结果与合成的siRNA结果一致。结论靶向OPNshRNA表达载体的构建成功为研究降低卵巢癌OPN表达的抗肿瘤效应,及与放化疗的协同效应打下了基础。

Construction and identification of targeting osteopontin shRNA expression vector

Objective To explore the new gene therapy for ovary cancer by the construction of osteopontin shRNA expression vector.Methods On the basis of OPN-mRNA genetic sequence database,an oligonucleotide chain consisting of 64 bases restriction sites at both ends was designed and synthesized.The annealed oligonucleotide chain was connected to the linearized plasmid pSilence with T4DNA ligase and was identified by enzyme digestion of recombinant plasmid and sequencing.Result The OPNsiRNA express vector cloning was s...