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靶向加工番茄elF4E1的CRISPR/Cas9载体有效性验证 被引量:1
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作者 付紫梅 袁伦 +3 位作者 邵冬南 郝小军 崔百明 向本春 《新疆农业科学》 CAS CSCD 北大核心 2018年第2期230-237,共8页
【目的】验证基于CRISPR/Cas9系统构建的靶向编辑加工番茄(Solanum lycopersicum)e IF4E1基因载体的有效性,为CRISPR/Cas9系统在培育PVY抗性植株中的应用提供技术支持。【方法】构建靶向编辑番茄真核翻译起始因子el F4E1基因的CRISPR/C... 【目的】验证基于CRISPR/Cas9系统构建的靶向编辑加工番茄(Solanum lycopersicum)e IF4E1基因载体的有效性,为CRISPR/Cas9系统在培育PVY抗性植株中的应用提供技术支持。【方法】构建靶向编辑番茄真核翻译起始因子el F4E1基因的CRISPR/Cas9系统表达载体,用农杆菌渗透法瞬时转化番茄植株,PCR扩增已转化植株靶位点周围DNA序列后用HaeⅢ进行酶切,回收未切开的条带与p GEM-T载体连接后进行单克隆测序。【结果】对测得的9个克隆序列进行比对分析,在PAM(protospacer adjacent motifs)上游的6~8 bp的碱基处均发生突变,并且都为单碱基的替换,导致多肽链中单个氨基酸的替换。【结论】利用CRISPR/Cas9基因组编辑系统构建的载体能够特异性地靶向加工番茄e IF4E1基因,为利用CRISPR/Cas9系统敲除e IF4E1基因,获得抗PVY病毒的番茄育种材料奠定了基础。 展开更多
关键词 加工番茄 PVY elf4e1 CRISPR/Cas9 瞬时转化
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The RALF1-FERONIA Complex Phosphorylates eIF4E1 to Promote Protein Synthesis and Polar Root Hair Growth 被引量:10
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作者 Sirui Zhu Jose Manuel Estevez +9 位作者 Hongdong Liao Yonghua Zhu Tao Yang Chiyu Li Yichuan Wang Lan Li Xuanming Liu Javier Martinez Pacheco Hongwei Guo Feng Yu 《Molecular Plant》 SCIE CAS CSCD 2020年第5期698-716,共19页
The molecular links between extracellular signals and the regulation of localized protein synthesis in plant cells are poorly understood.Here,we show that in Arabidopsis thaliana,the extracellular peptide RALF1 and it... The molecular links between extracellular signals and the regulation of localized protein synthesis in plant cells are poorly understood.Here,we show that in Arabidopsis thaliana,the extracellular peptide RALF1 and its receptor,the FERONIA receptor kinase,promote root hair(RH)tip growth by modulating protein synthesis.We found that RALF1 promotes FERONIA-mediated phosphorylation of elF4E1,a eukaryotic translation initiation factor that plays a crucial role in the control of mRNA translation rate.Phosphorylated elF4E1 increases mRNA affinity and modulates mRNA translation and,thus,protein synthesis.The mRNAs targeted by the RALF1-FERONIA-elF4E1 module include ROP2 and RSL4,which are important regulators of RH cell polarity and growth.RALF1 and FERONIA are expressed in a polar manner in RHs,which facilitate elF4E1 polar丨ocalization and thus may control local f?OP2 translation.Moreover,we demonstrated that high-level accumulation of RSL4 exerts negative-feedback regulation of RALF1 expression by directly binding the RALF1 gene promoter,determining the final RH size.Our study reveals that the link between RALF1-FERONIA signaling and protein synthesis constitutes a novel component regulating cell expansion in these polar growing cells. 展开更多
关键词 elf4e1 FER polar cell growth RALF1 root hairs RSL4
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