重组质粒pSUPER-HBs RNAi的构建及筛选
Journal Title: Di-er Junyi Daxue Xuebao - Year 2008, Vol 29, Issue 12
Abstract
目的:利用pSUPER-RNAi载体系统进行HBs RNAi序列的初步筛选和干扰效果鉴定。方法:设计并合成针对HBs基因区的3条siRNA(HBs-siRNA1、HBs-siRNA2、HBs-siRNA3),构建3种重组载体pSUPER-HBs-siRNA,与HBV质粒同时瞬转人胚肾293T细胞,72 h后观察转染效果,实时定量PCR和ELISA法鉴定3种干扰序列对HBs的干扰效果,筛选最佳干扰序列。结果:成功构建3种干扰质粒pSUPER-HBs-siRNA,能高效转染人胚肾上皮细胞293T,转染效率在70％以上；实时定量PCR和ELISA法结果均显示HBs-siRNA2可有效抑制HBs的表达,抵制率可达80%,而HBs-siRNA1、HBs-siRNA3干扰效果不显著,差异具有统计学意义(P<0.05)。结论:成功构建3种干扰质粒pSUPER-HBs-siRNA,筛选出其中能有效抑制HBs基因表达的pSUPER-HBs-siRNA2序列,为后续研究奠定了基础。
Authors and Affiliations
Xiang-ji LUO
Keywords
Related Articles
- EP ID EP82959
- DOI -
- Views 107
- Downloads 0
熵指数在老年患者七氟醚或丙泊酚靶控输注诱导中的应用
目的:探讨熵指数在老年患者七氟醚吸入或丙泊酚靶控输注诱导时的应用价值。方法:拟行气管插管全麻的老年患者40例,ASA Ⅰ~Ⅱ级,年龄≥65岁,随机分为七氟醚组(S组)和丙泊酚组(P组),分别以七氟醚吸入和丙泊酚靶控输注诱导,诱导期间调节熵指数维...
基于数据包络分析方法的某市民营医院服务效率实证研究
目的:应用数据包络分析(DEA)方法评估某市民营医院的服务效率,分析某市民营医院发展过程中经营战略存在问题。方法:采用问卷调查统计某市全部民营医院业务经营数据,筛选指标与样本,应用DEA方法对58家民营医院的服务效率进行评估。结果:经过...
Assessing mandibular bone mineral density with morphometric analysis
Objective：To determine the effectiveness and validity of morphometric analysis made from dental panoramic tomograms (DPTs) for predicting mandibular bone mineral density (BMD) while taking the result of dual energy Xra...
Immune tolerance of dendritic cells and its role in graft rejection
As an antigen-presenting cell, dendritic cells (DCs)have drawn great attention due to its powerful antigen-presenting ability and immunological regulation function. Recently, it is believed that DCs also participate in r...
Influence of delay in blood-processing on the level of cell-free fetal DNA in maternal plasma sample
Objective:To investigate the influencing factors of cell-free fetal DNA level in the maternal plasma during blood-processing.Methods: Aliquots of blood samples from pregnant women with male fetus were processed at 6 h an...