小鼠TRAF6特异性shRNA慢病毒质粒的构建及活性

目的:构建小鼠TRAF6基因特异性shRNA慢病毒质粒.方法:设计、合成小鼠TRAF6基因特异性shRNA核苷酸,将其链接入pENTR/U6载体,基因测序;通过LR克隆酶将pENTR/U6中的TRAF6特异性shRNA核苷酸插入CS -RfA -EG慢病毒载体,KpnⅠ限制酶切分析;通过慢病毒将TRAF6基因特异性shRNA慢病毒质粒导入MEF13细胞中,FACS分析质粒导入率,同时利用Western blot技术检测MEF13细胞中TRAF6的表达以及磷酸化IκBα的表达.以LacZ基因特异性shRNA慢病毒质粒为对照.结果:构建的小鼠TRAF6基因特异性shRNA慢病毒质粒与设计相符;MEF13细胞中慢病毒质粒的导入率为95％以上;与对照组比较,TRAF6基因特异性shRNA可长期、完全抑制MEF13细胞中TRAF6的表达及TRAF6介导的IκBα的磷酸化.结论:成功构建小鼠TRAF6基因特异性shRNA慢病毒质粒.     

Do Sprouting Tree Species on Erosion-prone Sites Carry Large Reserves of Resources?

Saplings ofEuptelea polyandra were studied to determine whethertree species found on unstable hillslopes of temperate, old-growthforests in Japan carry substantial storage materials for sproutingreplacement genets, as is the case with resprouter species offire-prone areas. Concentrations (% d. wt basis) of carbohydrates(starch, sucrose, glucose and fructose) contained in roots,stems and leaves were measured in summer and winter.E. polyandrasaplings were compared with those ofQuercus serrata (a frequentlysprouting tree), and those ofMallotus japonicus andIdesia polycarpa(rarely sprouting trees) in the same forest. Total concentrationsof carbohydrates (the sum of starch, sucrose, glucose and fructose)in roots were lowest inE. polyandra in both summer and winter.In addition,E. polyandra had a lower ratio of root biomass tototal plant biomass thanQ. serrata , but similar to that ofthe non-sprouting species,M. japonicus andI. polycarpa . Onthe other hand, the total concentration of carbohydrates inthe above-ground parts were similar in the four species in bothsummer and winter. These results indicate thatE. polyandra hadless long-term storage resources to implement sprouting, inspite of its apparent effectiveness in sprouting. We proposehypotheses to explain the reason whyE. polyandra stores a relativelysmall amount of resources for sprouting. Carbohydrate concentration; Euptelea polyandra Sieb. et Zacc; ground-surface disturbance; Idesia polycarpa Maxim; Mallotus japonicus (Thunb.) Muell. Arg.; Quercus serrata Thunb.; resprouter; root dry weight ratio; soluble sugars; sprouting; starch