稻瘟病菌热激蛋白复合体MoSsbl-MoSszl-MoZuol及SNARE蛋白MoSec22的互作蛋白在致病过程中的功能
发布时间:2022-01-26 04:36
由稻瘟病菌引起的稻瘟病是水稻上的三大病害之一,在世界各地均有发生,发生后造成不同程度的减产,给水稻等谷类作物的安全生产带来了严重威胁。目前防治稻瘟病主要采用选育抗病高产的优质品种,同时结合化学药物防治。由于稻瘟病菌生理小种易变,抗病品种种植过一段时间后其抗性容易丢失,所以防治稻瘟病一直是水稻种植生产上的一大难题。2005年稻瘟病菌全基因组测序完成,为从分子水平上对稻瘟病菌中的致病相关基因的功能和分子机制的研究提供了重要基础。本文主要有两部分内容:一部分是鉴定到了3个核糖体相关的分子伴侣蛋白,对它们在稻瘟病菌致病过程中功能进行分析,并解析其作用机制;另一部分是对实验室前期研究过SNARE蛋白MoSec22的互作蛋白进行筛选和鉴定,以求进一步探究其作用机制。分子伴侣是能够结合与稳定另一种蛋白质的构象,并通过有序地结合和释放,促进新生多肽链的折叠、多聚体的组装或降解,以及协助细胞器蛋白的跨膜运输的一类蛋白质,其中最大的一类为热激蛋白。本实验室前期发现蛋白激酶MoMkk1介导的MAPK信号途径控制稻瘟病菌的细胞壁完整性与致病力,为了进一步揭示其作用机制,我们采用co-IP的方法筛选MoMkk1...
【文章来源】:南京农业大学江苏省 211工程院校 教育部直属院校
【文章页数】:141 页
【学位级别】:博士
【部分图文】:
图2-2?SNARE蛋白在胞内膜转运过程中的功能(jahn?and?Scheller,2006)??Fig.?2-2?The?assignment?of?SNAREs?to?
稻瘟病菌热激蛋白复合体MoSsbl-MoSszl-MoZuol及SNARE蛋白MoSec22的互作蛋白在致病过程中的功能分析??Input?IP?Input?IP?Input?IP??-is::?iii:?a:??Ant卜S,??48?kD?—^?蓬涵—63?kD?.厚遍【漏—48?kD??图1-5?MoSsbl、MoSszl和MoZuol两两相互作用??Fig.?1-5?MoSsbl,?MoSszl?and?MoZuol?interact?with?each?other??Co-immunoprecipitation?assays?among?MoSsbl,?MoSszl?and?MoZuol.?The?following?strains?were??grown?in?liquid?CM?for?48?h:?1.?Guyl?1?containing?GFP?and?MoZuol-S;?2.?Guyl?1?containing??MoSsbl-GFP?and?MoZuol-S;?3.?Guyl?1?containing?GFP?and?MoSszl-S;?4.?Guyl?1?containing??MoSsbl-GFP?and?MoSszl-S;?5.?Guyl?1?containing?MoSszl-GFP?and?MoZuol-S.?Total?proteins??extracted?from?the?strains?were?subjected?to?SDS-PAGE?and?immunoblots?were?incubated?with?anti-GFP??and?anti-S?antibodies?as?indic
-Leu-Trp?SD-?H?is-Leu-T?rp-Ade??C?AD?AD-01688?AD?AD-01688?D?八?D?八?Din36i〕1?AD?AD-06601??ob?um?m?rm??Positive?Negative?Positive?Negative?Positive?Negative?Positive?Negative??SD-Leu-Trp?SD-His-Leu-Trp-Ade?SD-Leu-Trp?SD-His-Leu-Trp-Ade??图2-1酵母双杂交验证四个蛋白与MoSec22互作??Fig.?2-1?Verification?of?4?interacting-proteins?of?MoSec22?by?yeast?two-hybrid??Yeast?two?hybrid?assay?for?the?interaction?among?MoSec22?and?putative?proteins?identified?by?screening??cDNA?library?of?M.?oryzae.?The?cDNA?sequences?of?MoSEC22?were?inserted?into?the?vector?pGBKT7??and?the?cDNA?sequences?of?MGG一00588,?MGG_05202,?MGG—01688?and?MGG—06601?were?inserted??into?the?vector?pGADT7.?And?then?the?plasmids?were?co-transformed?into?yeast?AH?109?cells,?a
本文编号:3609827
【文章来源】:南京农业大学江苏省 211工程院校 教育部直属院校
【文章页数】:141 页
【学位级别】:博士
【部分图文】:
图2-2?SNARE蛋白在胞内膜转运过程中的功能(jahn?and?Scheller,2006)??Fig.?2-2?The?assignment?of?SNAREs?to?
稻瘟病菌热激蛋白复合体MoSsbl-MoSszl-MoZuol及SNARE蛋白MoSec22的互作蛋白在致病过程中的功能分析??Input?IP?Input?IP?Input?IP??-is::?iii:?a:??Ant卜S,??48?kD?—^?蓬涵—63?kD?.厚遍【漏—48?kD??图1-5?MoSsbl、MoSszl和MoZuol两两相互作用??Fig.?1-5?MoSsbl,?MoSszl?and?MoZuol?interact?with?each?other??Co-immunoprecipitation?assays?among?MoSsbl,?MoSszl?and?MoZuol.?The?following?strains?were??grown?in?liquid?CM?for?48?h:?1.?Guyl?1?containing?GFP?and?MoZuol-S;?2.?Guyl?1?containing??MoSsbl-GFP?and?MoZuol-S;?3.?Guyl?1?containing?GFP?and?MoSszl-S;?4.?Guyl?1?containing??MoSsbl-GFP?and?MoSszl-S;?5.?Guyl?1?containing?MoSszl-GFP?and?MoZuol-S.?Total?proteins??extracted?from?the?strains?were?subjected?to?SDS-PAGE?and?immunoblots?were?incubated?with?anti-GFP??and?anti-S?antibodies?as?indic
-Leu-Trp?SD-?H?is-Leu-T?rp-Ade??C?AD?AD-01688?AD?AD-01688?D?八?D?八?Din36i〕1?AD?AD-06601??ob?um?m?rm??Positive?Negative?Positive?Negative?Positive?Negative?Positive?Negative??SD-Leu-Trp?SD-His-Leu-Trp-Ade?SD-Leu-Trp?SD-His-Leu-Trp-Ade??图2-1酵母双杂交验证四个蛋白与MoSec22互作??Fig.?2-1?Verification?of?4?interacting-proteins?of?MoSec22?by?yeast?two-hybrid??Yeast?two?hybrid?assay?for?the?interaction?among?MoSec22?and?putative?proteins?identified?by?screening??cDNA?library?of?M.?oryzae.?The?cDNA?sequences?of?MoSEC22?were?inserted?into?the?vector?pGBKT7??and?the?cDNA?sequences?of?MGG一00588,?MGG_05202,?MGG—01688?and?MGG—06601?were?inserted??into?the?vector?pGADT7.?And?then?the?plasmids?were?co-transformed?into?yeast?AH?109?cells,?a
本文编号:3609827
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