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玉米YL蛋白质与ZmPEPCs互作及对其酶活性的影响

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玉米YL蛋白质与ZmPEPCs互作及对其酶活性的影响

作者：肖乐君华中农业大学

学位级别：硕士

Maize（Zea mays L.）is one of the important crops,which plays an important role in national economy and biological *** are important materials for studying plant growth and *** this study,candidate gene GRMZM2G095254（yl gene）of yellow leaf mutant（yellow leaf,yl）and Weaky mutant（Weaky）was *** on the interaction between YL protein and CPEPC,CPEPC was previously verified in the *** study explored the interaction between Root-PEPC and YL protein,and studied the influence of the interaction between YL protein and Zm PEPCs on their enzymatic *** main results are as follows:*** the amino acid sequences of CPEPC,CPEPC,Root-PEPC and Bacterial-PEPC in maize,it was found that the amino acid sequences of the three plant type PEPCs have high homology,but the amino acid sequences of plant type PEPCs and bacterial type PEPCs have low homology,and there are significant differences in gene *** structural differences of the four PEPCs amino acid sequences may be related to the differences in the catalytic and regulatory properties of their *** two-hybrid and BiFC experiments have shown that there is interaction between Root-PEPC and YL protein,with interaction sites located in the nucleus and weak interaction signals on the cell *** protein,CPEPC,CPEPC and Root-PEPC were expressed in Escherichia *** enzyme activity of Zm PEPCs was measured at different temperatures with YL *** results showed that the enzyme activities of CPEPC,CPEPC and Root-PEPC were all highest at 30℃.After adding YL protein and ATP simultaneously to the PEPC enzyme activity measurement system,the enzyme activity of Zm PEPCs was significantly decreased under various temperature *** protein can inhibited the enzyme activity of Zm PEPCs,and the inhibitory effect of YL protein was an active process,and its activation required ATP *** the PEPC enzyme activity of the leaves in the two-le

关键词： Maize yl gene yeast two-hybrid BiFC PEPC Prokaryotic expression Enzymatic properties

Screening of human vascular endothelial growth factor (VEGF) receptor Flt-1 domain and study on its biological activity

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Science China(Life Sciences) 2001年第5期44卷 497-505页

作者：马骊张智清王小宁孙大军周小明陈爱君姚立红 Institute of Molecular Immunology The First Military Medical UniversityGuangzhou 510515China

Four human vascular endothelial growth factor receptor Flt-1 cDNA fragments containing extracellular domain loops 2,1-2,2-3 and 1-3 respectively were amplified from human placental cDNA library by PCR and used for screening ligand binding domains by yeast two-hybrid *** result showed that,not only loop 1-3,but also the smaller fragment loop 2-3 could bind bo *** expression plasmids pPIC9K/Flt-1(1-3)and pPIC9K/Flt-1(2-3)were constructed and transformed to *** host strain GS115,cultured in flasks,and expressed under the induction of 1% *** expressed product existed in supernatant in the form of soluble molecules and contained more than 60% of total protein after being induced for *** being purified by CM-Sepharose FF and Sephacryl S-100 chromatography,its purity reached above 90%.Biological assay in vitro showed that the binding capacity of expressed soluble Flt-1(2-3)to hVEGF165 and its nhibiting effect on the proliferation of human umbilical reins endothelial cells(HUVEC)stimulated with hVEGF165 were close to those of s Flt-1(1-3).Animal test showed that sFlt-1(2-3)could nhibit the formation of regenerate blood vessels stimulated with hVEGF165 significantly.

关键词： vascular endothelial growth factor (VEGF) receptor yeast two-hybrid Pichia. pastoris gene expression.

Cloning of Rice Blast Resistance Gene Pik_2-H4 and Screening of Its Interaction Protein

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Plant Diseases and Pests 2014年第4期5卷 26-30页

作者： Liu Hao Sun Dayuan Wang Hui Chen Zhiqiang National Engineering Research Center of Plant Space Breeding South China Agricultural University

Abstract The broad-spectrum rice blast resistance gene Pik2-H4 was cloned from the highly resistant line H4. The full-length sequence of Pik2-H4 was 3 066 bp, which was different from the corresponding allele sequence of Nipponbare. Pik2-H4 belonged to NBS-LRR genes and coded a protein containing NB-ARC domain and leueine-rieh repeats. To find the interaction proteins with Pik2-H4 from the rice, the yeast two-hybrid system was used and three important proteins （ LOC- Os08939300, LOCOs03g25960 and LOC-Os09929130）were identified. The results could provide some new information for the mechanism of rice blast resistance mediated by Pik2-H4.

关键词： Pik2-H4 Clone yeast two-hybrid Interaction protein

Interactions between Toxoplasma gondii micronemal protein MIC3 and host proteins Spata3 and Dkk2

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Interactions between Toxoplasma gondii micronemal protein MI...

中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会

作者： Yifan Wang Rui Fang Yuan Yuan Ming Pan Min Hu Yanqin Zhou Bang Shen Junlong Zhao State Key Laboratory of Agricultural Microbiology College of Veterinary MedicineHuazhong Agricultural University Key Laboratory of development of veterinary diagnostic products Ministry of AgricultureHuazhong Agricultural University

As an obligate intracellular protozoan,Toxoplasma gondii is a successful pathogen infecting a variety of animals and *** host infection,micronemal proteins(MICs) are released to parasite surface,where they bind host cells to mediate gliding motility and host-cell *** an adhesin,the micronemal protein MIC3 plays important roles in host cell attachment,as well as modulation of host EGFR signaling ***,the specific host proteins that interact with MIC3 are unknown and the identification of such proteins will increase our understanding of how MIC3 exerts its *** this study,we sought to identify host proteins interacting with MIC3 by yeast two-hybrid *** MIC3 as bait,we screened a library expressing mouse proteins and found five mouse proteins that showed positive interactions with *** of which,spermatogenesis-associated protein 3(Spata3) and dickkopf-related protein 2(Dkk2),were further confirmed to interact with MIC3 both in vitro and in vivo by additional protein-protein interaction *** also found that the tandem repeat EGF domains of MIC3 were critical in mediating the interactions with the identified host *** is the first study to show that MIC3 interacts with host proteins that are involved in reproduction,growth and *** results will help us to better understand the functions of adhesion-associated micronemal proteins in Toxoplasma gondii.

关键词： Toxoplasma gondii MIC3 yeast two-hybrid Dkk2 Spata3

Screen of host proteins interacting with Toxoplasma gondii rhoptry protein ROP9

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Screen of host proteins interacting with Toxoplasma gondii r...

中国动物学会寄生虫学专业委员会第十届全国寄生虫学青年工...

作者： Zexiang Wang Dongmei Yue Siyang Huang Dezhen Gao Xinquan Zhu State Key Laboratory of Veterinary Etiological Biology Key laboratory of Veterinary Parasitology of Gansu Province Lanzhou Veterinary Research Institute Chinese Academy of Agricultural Sciences College of Animal Science and Veterinary Medicine Heilongjiang Bayi Agricultural University

Toxoplasma gondii is the caustive agent of toxoplasmosis and can infect almost any warm-blooded animals, including human. Rhoptry protein 9(ROP9) is a 36 kDa soluble protein present only in the tachyzoite stage during *** life cycle and participate in the early stages of parasite invasion. Until up to now, host proteins interacting with ROP9 still have not been found. In our study, a yeast two-hybrid system was used to screen host proteins interacting with ROP9. ROP9 was inserted into the pGBKT7 vector as a bait which expressed fusion protein with the GAL4 DNA-binding domain. Bait proteins expression detection was carried on via western blotting and the autoactivation of bait plasmid was tested compared with the empty vector pGBKT7. A human brain c DNA library was used to identified host proteins interacting with ROP9 by yeast-two-hybrid system. After screening, positive hits were sequenced and analysed with BLAST analysis. We obtained 5 host protein sequences and four of which are false positives. The interaction between ROP9 and cytochrome coxidase11(COX11) needs to be further confirmed. Our study firstly report interactions between Toxoplasma gondii ROP9 and host proteins. The results will facilitate the understanding of the function of ROP9 and host signal tranduction ROP9 involved.

关键词： Toxoplasma gondii ROP9 Interaction yeast two-hybrid COX11

A conserved Puccinia striiformis effector interacts with wheat NPR1 and reduces induction of pathogenesis-relatedgenes in response to pathogens

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A conserved Puccinia striiformis effector interacts with whe...

中国植物病理学会2016年学术年会

作者： WANG Xiao-dong YANG Bao-ju LI Kun KANG Zhen-sheng Dario Cantu Jorge Dubcovsky Department of Plant Science University of California State Key Laboratory of Crop Stress Biology for Arid Areas and College of Plant Protection Northwest Agriculture and Forestry University Department of Plant Pathology Agriculture University of Hebei Department of Viticulture and Enology University of CaliforniaDavisCA 95616USA.5.Howard Hughes Medical Institute(HHMI)Chevy Chase

In Arabidopsis,NPR1 is a key transcriptional co-regulator of systemic acquired *** pathogen challenge,NPR1 translocates from the cytoplasm to the nucleus,where it interacts with TGA-bZIP transcription factors to activate the expression of several pathogenesis-related *** a screen of a yeast two-hybrid library from wheat leaves infected with ***,we identified a conserved rust effector that interacts with wheat NPR1 and named it Puccinia NPR1 interactor（PNPi）.PNPi interacts with the NPR1/NIM1-like domain of NPR1 via its C-terminal *** bimolecular fluorescence complementation assays,we detected the interaction between PNPi and wheat NPR1 in the nucleus of Nicotiana benthamiana protoplasts.A yeast three-hybrid assay showed that PNPi interaction with NPR1 competes with the interaction between wheat NPR1 and *** barley transgenic lines over expressing PNPi,we observed reduced induction of multiple PR genes in the region adjacent to Pseudomonas *** DC3000 *** on these results,we hypothesize that PNPi has a role in manipulating wheat defense response via its interactions with NPR1.

关键词： Wheat Stripe rust NPR1 Pathogen effector yeast two-hybrid Bimolecular fluorescence complementation Transgenic barley

Preliminary identification of the function of Ralstonia solanacearum effector protein Rip75 and screening of candidate interaction proteins

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Preliminary identification of the function of Ralstonia sola...

中国植物病理学会2021年学术年会

作者： HEN Kun LI Huaqi GAO Meijia WANG Shanshan DANG Hao WEI Jiaxian ZHUANG Weijian College of Plant Protection Fujian Agriculture and Forestry University College of Agronomy Fujian Agriculture and Forestry University

Bacterial wilt caused by Ralstonia solanacearum is harmful to many *** pathogen caused water shortage due to blocking effect after the propagation of plant vascular bundle,which affected the normal growth and development of plants,and caused the wilt and death,resulting in crop production reduction and huge economic *** the process of pathogen infection,the bacterial use type Ⅲ secretion system to inject a large number of effector proteins directly into host cells,which may affect the immune response of plants and lead to the host plant infected or non-host plants to have hypersensitive *** is a complex species,which can secrete more than 100 kinds of effector proteins,but the function of most of them is not *** whole genome of *** has been sequenced in this laboratory,and 75 effector proteins were predicted by T3 Es *** this study,we selected Rip75 as candidate effector protein gene to clone and preliminarily analyze the ***75,which encodes a total of 338 amino acids and has no conserved domain,can cause hypersensitive response in Nicotiana *** is a new effector protein that has not been reported in *** the mutant strain of Δrip75 was constructed by homologous *** inoculating peanut varieties resistant and susceptible to bacterial wilt,it was found that the pathogenicity of mutant strain Δrip75 to peanut was *** proteins（corresponding to AhPRPS,AhKUAl and AhSBT） interacting with Rip75 were screened by yeast two-hybrid method,and their interaction was verified by BIFC *** results preliminarily revealed the function of the effector protein Rip75 and speculated that it may be a toxicprotein,which provided a theoretical basis for the resistance and control of peanut bacterial wilt.

关键词： Ralstonia solanacearum Peanut Rip75 Effector yeast two-hybrid

Identification of an interaction between calcium-dependent protein kinase 4(EtCDPK4) and serine protease inhibitor(EtSerpin) in Eimeria tenella

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Identification of an interaction between calcium-dependent p...

第十一届全国寄生虫学青年工作者学术研讨会

作者： Ling Lv Huanzhi Zhao Bing Huang Qiping Zhao Hui Dong Shunhai Zhu Zhihang Li Lu Wang 韩红玉 Shanghai Veterinary Research Institute Chinese Academy of Agricultural SciencesKey Laboratory of Animal Parasitology of Ministry of Agriculture College of Life and Environment Sciences Shanghai Normal University

Background:Eimeria tenella is an obligate intracellular apicomplexan protozoan parasite that has a complex *** ions,through various calcium-dependent protein kinases(CDPKs),regulate key events in parasite growth and development,including protein secretion,movement,differentiation,and invasion of and escape from host *** this study,we identified proteins that interact with Et CDPK4 to lay a foundation for clarifying the role of CDPKs in calcium ***:Eimeria tenella merozoites were collected to construct a yeast two-hybrid(Y2 H) c DNA *** Y2 H system was used to identify proteins that interact with Et *** of interacting proteins was confirmed using bimolecular fluorescence complementation and co-immunoprecipitation in ***-localization of proteins was performed using immunofluorescence ***:Eight proteins that interact with Et CDPK4 were identified using the Y2 H *** of the proteins,*** serine protease inhibitor 1(Et Serpin),was further ***:In this study,we screened for proteins that interact with Et *** interaction between Et Serpin and Et CDPK4 was identified that may contribute to the invasion and development of *** in host cells.

关键词： Eimeria tenella Calcium-dependent protein kinases Serpin yeast two-hybrid Bimolecular fluorescence complementation Co-immunoprecipitation

Lescpth5诱饵载体的构建及其自激活作用的检测(英文)

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Plant Diseases and Pests 2011年第4期2卷 1-2,5页

作者：张月娟万一陈华民张琨赵廷昌陕西省微生物研究所陕西西安710043 中国农业科学院植物保护研究所北京100193

[ Objective ] The paper was to construct bait vector pGBKTT-Lescpth5, and detect its self-activated transcription activity and the toxicity on yeast cells. [ Method] Lescpth5 was amplified by PCR technique, and connected into bait vector pGBKTT. The recombined vector was transformed into yeast competent ceils AH109 and carried out self-activated detection and toxicity detection in the *** medium. [ Result] Digestion and sequencing results showed that bait vector pGBKT7-Lescpth5 was suecessfully constructed with correct reading frame. Self-activated activity and toxicity detection results showed that bait vector had no self- activated activity on yeast strain AH109, which also had no toxicity on yeast. [Condusion] Bait vector successfully constructed could be used in yeast two-hybrid system, which laid the foundation for screening of cDNA library in the next step.

关键词： Pseudomonas syringae pv. tomato Bait vector yeast two-hybrid Self-activated China