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集思慧遠客戶發表《耶氏解脂酵母Y-2對赭曲霉毒素A降解反應的蛋白質和轉錄組分析》

幸运飞艇官方开户FABURIQI:2019-06-28 LIULANCISHU:363

HEZUODANWEI:JIANGSUDAXUESHIPINYUSHENGWUGONGCHENGXUEYUAN

FABIAOQIKAN:Annals of Applied Biology

YINGXIANGYINZI:IF=2.046

研究背景

      ZAIGUWU、SILIAOHESHIPINZHONG,CENGBAODAOGUOochratoxin A(OTA)DEFASHENG。OTADESHENGWUFANGZHIBEIRENWEISHIYIZHONGHENYOUQIANTUDECELVE。XIANQIANYANJIUBIAOMINGYESHIJIEZHIJIAOMUY-2KEJIANGJIEOTA。BENYANJIUCAIYONGDANBAIZHIZUXUEHEZHUANLUZUFENXIDEFANGFA,YANJIULEYESHIJIEZHIJIAOMUY-2JIANGJIEOTADEKENENGJIZHIJIQIJIEKANGJIAOMUDEFANYING。JIEGUOBIAOMING:YESHIJIEZHIJIAOMUY-2ZHONGCANYUQUCHUHUOXINGO2YOUGUANDEDANBAIZHIHEJIYINJINXINGLESHANGDIAO,YIXIANGYINGOTAYINQIDEYANGHUAYINGJI。CIWAI,ZAIYINGJITIAOJIANXIACHUQUCHANSHENGDEDUWUSUOSHEJIDEYIXIEDANBAIZHIHEJIYINBEISHANGDIAO,YIGAISHANZHEZHONGJIEKANGJIAOMUDUIOTAYALIDENAISHOUXING。CIWAI,YIXIEBIANMAZHUANYUNDANBAIHEDUOYAONAIYAODANBAIDEJIYINBEIXIANZHUSHANGDIAO,CONGERCONGXIBAOZHONGZHUANYUNHEPAICHUOTA,JIANQINGDUIYESHIJIEZHIJIAOMUY-2DEDUXING。ZHIDEZHUYIDESHI,ZHIJIECANYUOTAJIANGJIEDEECM14JIYIN(BIANMAJINSHUSUOTAIMEIECM14)ZAIGAIZHENJUNDUSUCHULIDEY-2ZHONGBIAODASHANGDIAO。WEILEDIYUOTAXIEPO,OTAYOUDAOLEYIXIEYUSANSUOSUANXUNHUANXIANGGUANDEJIYIN,WEIYESHIJIEZHIJIAOMUY-2CHANSHENGFENGFUDENENGLIANG。CIWAI,CANYUDNAFUZHIHEXIUFUDEYIXIEJIYINDEXIADIAOYEJIANGDILEYESHIJIEZHIJIAOMUY-2XIUFUOTASUOZHIDNASUNSHANGDENENGLI。ZHEXIEJIEGUOKENENGYIWEIZHUZHEZHONGJIEKANGJIAOMUDEOTAJIANGJIESHIDUOZHONGFANGYUFANYINGDUIOTAYINGLIDEXIETONGZUOYONG。

材料與方法

DANBAIZUFENXI

幸运飞艇官方开户 YANGPINYONGLIANG:1×108cell/mL;

幸运飞艇官方开户 JIANCEPINGTAI:2DNINGJIAODIANYONG;JIZHIFUZHUJIGUANGJIEXI/DIANLIFEIXINGSHIJIANZHIPU(MALDI-TOF/MS);

CHAYIDANBAISHAIXUAN:fold change≥1.5 or ≤0.67QIEp<0.05;

ZHUANLUZUFENXI

CEXUPINGTAI:Illumina Hiseq 2500 PE150;

CHAYIJIYINSHAIXUAN:log2 fold change >1 (FDR < 0.05)QIEp<0.05;

幸运飞艇官方开户 FENXINEIRONG:CHAYIJIYINGOGONGNENGZHUSHIJIKEGGsGONGNENGZHUSHI;

RT-qPCRFENXI

幸运飞艇官方开户 XUANQU16GEJIYINJINXINGBIAODALIANGSHUIPINGDEYANZHENG;


主要研究結果

01 蛋白組分析

幸运飞艇官方开户       YONG2-DEFENLIDEOTAHUOBUJINGOTACHULIDEYESHIJIEZHIJIAOMUY-2DESUOYOUDANBAIZHIRUTU1a,bSUOSHI。HULVEFEICHANGWEIRUODEBANDIANHEBUQUEDINGXINGZHUANGHEQUYUDEBANDIAN,ZAIMEIGENINGJIAOZHONGGONGJIANCEDAO512GEDANBAIZHIDIAN,QIZHONG77GEDEPs(PINGJUNfold change≥1.5HUO≤0.67,p<0.05)。CIWAI,TONGGUOMALDI-TOF/MSJIANDING66GEDEPs,BAOKUO43GESHANGDIAODEDANBAIHE23GEXIADIAODANBAI。

圖1不含(A)和含1μg/mL OTA(B)處理的耶氏解脂酵母Y-2蛋白的2D凝膠電泳

圖2差異蛋白的功能分類

02 轉錄組分析

幸运飞艇官方开户       YONGRNA-SeqFENXILEJINGOTACHULIHUOBUJIAOTACHULIDEY-2DEZHUANLUZUFEN,BINGJIANGQICUNRUGEOSHUJUKU,DENGLUHAOWEIGSE123081。ZONGGONGHUODELEZHELIANGZHONGCHULIDE27,033,428HE22,439,334GEclean data,BINGDUIOTACHULIDEYESHIJIEZHIJIAOMUY-2HEBUJINGOTACHULIDEYESHIJIEZHIJIAOMUY-2DEJIYINBIAODASHUIPINGJINXINGLEBIJIAO。JIEGUOBIAOMING,1,156GEJIYINBIAODACHAYI,QIZHONG691GEJIYINBIAODASHANGDIAO(log2Y+OTA/Y>1,FDR<0.05),465GEJIYINBIAODAXIADIAO(log2Y+OTA/Y<?1,FDR<0.05)(TU3)。ZAIJINGOTACHULIDEYESHIJIEZHIJIAOMUY-2ZHONG,250GEJIYINMINGXIANSHANGDIAO(log2Y+OTA/Y>2,FDR<0.05),211TIAOJIYINXIANZHUXIADIAO(log2Y+OTA/Y<?2,FDR<0.05)。

圖3不含和含1μg/mL OTA處理的耶氏解脂酵母Y-2的差異表達基因


圖4差異表達基因的GO富集分析



圖5差異表達基因的KEGG富集分析

03 RT-qPCR分析

        CONGZHUANLUZUFENXIJIEGUOLAIKAN,CANYUYESHIJIEZHIJIAOMUY-2DUIOTAFANYINGDEJIYINBAOKUOAN5440,CCP1,yakc,AflN,gto2,MFS,CDR1,sdh1,sdh3,acob,icdA,ECM14,SGS1,pol3HErad15,WEILEYANZHENGZHEXIEJIYINDEBIAODASHUIPING,SHIYONGLEBIAO1SUOSHIDEYINWU。RT-qPCRJIEGUOXIANSHIOTACHULIHOUYESHIJIEZHIJIAOMUY-2DEAN5440(TU6a)、CCP1(TU6b)、yakc(TU6c)、AflN(TU6d)、gto2(TU6e)、MFS(TU6f)、CDR1(TU6g)、sdh1(TU6h)、sdh3(TU6i)、acob(TU6J)、icdA(TU6k)HEECM14(TU6l)DEXIANGDUIBIAODASHUIPINGJUNZENGJIA。CIWAI,YESHIJIEZHIJIAOMUY-2DESGS1(TU6m)、pol3(TU6n)HErad15(TU6o)BEIOTAXIADIAO。RT-qPCRDEBIAODAJIEGUOYUZHUANLUZUSHUJUYIZHI。

圖6用RT-qPCR方法驗證了耶氏解脂酵母Y-2對OTA的反應中不同表達的基因(部分)


總結

      DANBAIZHIZUHEZHUANLUZUFENXIDEJIEGUOBIAOMING,YESHIJIEZHIJIAOMUY-2DEFANGYUXITONGBEIJIHUO,YIXIANGYINGOTAYINQIDEYANGHUAYINGJI,ZAIYINGJITIAOJIANXIA,YIXIECANYUQINGCHUCHANSHENGDEDUWUDEDANBAIZHIHEJIYINYEBEISHANGDIAO。CIWAI,BIANMAZHUANYUNDANBAIDEXIANGGUANJIYINBEISHANGDIAO,CONGXIBAOZHONGZHUANYUNHEPAICHUOTA,CONGERJIANQINGLEYESHIJIEZHIJIAOMUDUIY-2DEDUXING。TONGSHI,ZHIJIECANYUOTAJIANGJIEDEJIYINECM14ZAIGAIZHENJUNDUSUCHULIDEYESHIJIEZHIJIAOMUY-2ZHONGBIAODASHANGDIAO。WEILEFANGYUOTAXIEPO,YIXIEDANBAIZHIHEBIANMAZAITCAXUNHUANZHONGQIZHONGYAOZUOYONGDEMEIDEJIYINBEISHANGDIAOYICHANSHENGJIEZHIYESHIJIAOMUY-2DECHONGZUNENGLIANG。CIWAI,CANYUDNAFUZHIHEXIUFUDEJIYINDEXIADIAOYEJIANGDILEYESHIJIEZHIJIAOMUY-2XIUFUOTASUOZHIDNASUNSHANGDENENGLI。ZHEXIEJIEGUOBIAOMING,GAIJIEKANGJIAOMUDUIOTADEJIANGJIEKENENGSHIDUIOTAXIEPODEDUOZHONGFANGYUFANYINGDEXIETONGXIAOYING。BENYANJIUYOUZHUYULEJIEYESHIJIEZHIJIAOMUY-2JIANGJIEOTADEFENZIJIZHIYIJIGAIJIEKANGJIAOMUDUIOTADEXIANGYING。


DIANJILIANJIE:http://onlinelibrary.wiley.com/doi/abs/10.1111/aab.12515CHAKANLUNWENQUANWEN。