Gravity-induced root curvature involves the asymmetric distribution of the phytohormone auxin.This response depends on the concerted activities of the auxin transporters such as PIN-FORMED(PIN)proteins for auxin efflu...Gravity-induced root curvature involves the asymmetric distribution of the phytohormone auxin.This response depends on the concerted activities of the auxin transporters such as PIN-FORMED(PIN)proteins for auxin efflux and AUXIN RESISTANT 1(AUX1)for auxin influx.However,how the auxin gradient is established remains elusive.Here we identified a new mutant with a short root,strong auxin distribution in the lateral root cap and an impaired gravitropic response.The causal gene encoded an Arabidopsis homolog of the human unconventional prefoldin RPB5 interactor(URI).At URI interacted with prefoldin 2(PFD2)and PFD6,twoβ-type PFD members that modulate actin and tubulin patterning in roots.The auxin reporter DR5_(rev):GFP showed that asymmetric auxin redistribution after gravistimulation is disordered in aturi-1 root tips.Treatment with the endomembrane protein trafficking inhibitor brefeldin A indicated that recycling of the auxin transporter PIN2 is disrupted in aturi-1 roots as well as in pfd mutants.We propose that At URI cooperates with PFDs to recycle PIN2and modulate auxin distribution.展开更多
文摘目的构建含rPB- DR启动子和TK自杀基因的重组复制缺陷型腺病毒并进行鉴定,为前列腺癌基因治疗的研究奠定基础。方法应用分子克隆技术,将rPB DR启动子连接至pBluescriptSK -TK上构建出pBluescriptSK -rPB- DR TK,再酶切出rPB -DR -TK目的片断,并将其连接至穿梭质粒pShuttle上。将构建好的pShuttle- rPB- DR -TK经PmeⅠ单切线性化后再以电穿孔法转化至E.coli.BJ5183AdEasy1内进行同源重组,抽提质粒经卡那抗性筛选及酶切鉴定正确的即为pAd- rPB- DR -TK,再经PacⅠ酶切回收后以脂质体法转染人胚肾293细胞,包装出完整腺病毒,然后大量扩增测定滴度,并行PCR鉴定。结果经酶切、PCR及测序鉴定证实成功构建了包含rPB -DR启动子和TK自杀基因的重组复制缺陷型腺病毒pAd rPB DR TK,包装、扩增后测病毒滴度为1.1×108TCID50/ml。结论成功构建出包含rPB DR启动子和TK自杀基因的重组复制缺陷型腺病毒并鉴定正确,可用于下一步前列腺癌基因治疗的研究中。
基金supported by an initiation fund from the Shandong Agricultural University,the National Natural Science Foundation of China(31570291)the Shandong“Foreign experts double hundred”Program(WST2017008)the Natural Science Foundation of Shandong Province(ZR2021MC175)。
文摘Gravity-induced root curvature involves the asymmetric distribution of the phytohormone auxin.This response depends on the concerted activities of the auxin transporters such as PIN-FORMED(PIN)proteins for auxin efflux and AUXIN RESISTANT 1(AUX1)for auxin influx.However,how the auxin gradient is established remains elusive.Here we identified a new mutant with a short root,strong auxin distribution in the lateral root cap and an impaired gravitropic response.The causal gene encoded an Arabidopsis homolog of the human unconventional prefoldin RPB5 interactor(URI).At URI interacted with prefoldin 2(PFD2)and PFD6,twoβ-type PFD members that modulate actin and tubulin patterning in roots.The auxin reporter DR5_(rev):GFP showed that asymmetric auxin redistribution after gravistimulation is disordered in aturi-1 root tips.Treatment with the endomembrane protein trafficking inhibitor brefeldin A indicated that recycling of the auxin transporter PIN2 is disrupted in aturi-1 roots as well as in pfd mutants.We propose that At URI cooperates with PFDs to recycle PIN2and modulate auxin distribution.
