【摘 要】
：
目的：构建人活性转录因子4(activating transcriptional facror4,ATF4)慢病毒载体并包装慢病毒颗粒(lentivirus),探讨成肌细胞C2C12中ATF4基因慢病毒修饰对BMP2诱导其成骨分化的作用.方法 设计并合成人ATF4基因的上、下游引物,采用PCR方法从真核质粒pcDNA3.1(-)-ATF4中扩增目的基因.鉴定后与慢病毒载体pWPT-GFPV5进行重
【机 构】
：
重庆医科大学基础医学院细胞生物学及遗传学教研室,重庆,400016
【出 处】
：
中国细胞生物学学会2015年全国学术大会
论文部分内容阅读
目的：构建人活性转录因子4(activating transcriptional facror4,ATF4)慢病毒载体并包装慢病毒颗粒(lentivirus),探讨成肌细胞C2C12中ATF4基因慢病毒修饰对BMP2诱导其成骨分化的作用.方法 设计并合成人ATF4基因的上、下游引物,采用PCR方法从真核质粒pcDNA3.1(-)-ATF4中扩增目的基因.鉴定后与慢病毒载体pWPT-GFPV5进行重组,再将重组慢病毒载体pWPT-GFPATF4与慢病毒包装质粒pMD2G、pSPAX2共转入293T细胞中,进一步包装成ATF4慢病毒(Lv-ATF4)；用病毒上清感染成肌细胞C2C12,观察绿色荧光蛋白(GFP)感染效率,并应用流式细胞仪(FCM)监测BMP2诱导成肌细胞C2C12细胞分化时对其增殖凋亡的影响,蛋白质印迹法监测凋亡相关基因Cleaved Caspase-3、Chop与p-JNK蛋白的表达,电镜观察细胞形态差异.
其他文献
Heart failure is an end stage disease which can be caused by various pathological insults, such as systemic hypertension, ischemic heart injuries, virus infections, and exposures to reagents with card
Hippo信号通路是果蝇以及哺乳动物当中调控器官大小的重要通路。被激活的Hpo通过磷酸化级联反应最终增加Yki的磷酸化，从而阻止Yki入核行使转录因子的功能。通过实验我们发现，当用c587-gal4在果蝇卵巢生殖细胞周围的护卫细胞(EC)中敲低Yki的表达，能够产生生殖细胞过度增殖、延缓分化的表型。我们同时敲低Yki以及其共同作用因子Sd后，能够使这种表型得到回复。所以说Yki的这种表型还是基于经
Tubular organs support the uptake and exchange of nutrition, metabolites, and gases that are essential for the normal physiology of animals.The notochord of ascidian Ciona forms a tube consisting of o
The inhibitory role of p53 in DNA double-strand break (DSB) repair seems contradictory to its tumor suppressing property.The p53 isoform Δ113p53/Δ133p53 is a p53 target gene that antagonizes p53 apopt
The zebrafish sensory posterior lateral line (pLL) has become an attractive model for studying collective cell migration and cell morphogenesis.Recent studies have indicated that chemokine, Wnt/β-cate
The serine/arginine-rich splicing actor 4 (SRSF4) is essential for pre-mRNA splicing and can influence alternative splice site choice, yet little is known about the specific function of this gene in r
The liver has high regenerative capacity, but it is not clear whether most biliary cells (particularly larger cholangiocytes) transdifferentiate into hepatocytes in regenerating liver.
PeCPD and PeDWF4 are separately the homologous to Arabidopsis CPD (AtCPD) and DWF4 (AtDWF4) isolated from Populus euphratica.
Mutations in Neurofibromin 1 (NF1) have been implied in multiple human diseases, including cancers and neurological disorders.We have observed that brain specific deletion of NF1 affected the balance
Protein arginine methyltransferase 6 (PRMT6), as a nucleus enzyme, mainly involves in transcriptional regulation, DNA repair, and histone modification.However, only few are reported about the biologic