全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

小麦ATG8的原核表达及其抗血清制备

DOI: 10.3969/j.issn.1000-7091.2013.01.019, PP. 101-105

Keywords: ATG8,免疫印迹,抗血清,融合蛋白,原核表达

Full-Text   Cite this paper   Add to My Lib

Abstract:

提取小麦品种L10的总RNA,进行反转录,其产物用于探究自噬基因ATG8的生物学功能。用PCR方法克隆ATG8,将克隆的ATG8亚克隆至表达载体PMD19-T,酶切后切胶回收,然后把产物测序鉴定,转化宿主菌E.coliRosetta-gamiB(DE3),构建原核表达系统,将融合蛋白纯化后制备其兔抗血清,然后利用WesteRnBlotting技术对兔抗血清进行检测,鉴定了该抗血清的结合特异性。ATG8抗体的成功制备,为小麦中ATG8基因和自噬功能的研究奠定了基础。

References

[1]  Nakatogawa H,Suzuik K,Kamada Y, et al. Dynames and diveRsity in autophagy mech anisms: lessons fRom yeast
[2]  [J]. Nat Rev Mol Cell Biol, 2009, 10( 7) : 458 - 467.
[3]  Bassham D C,LapoRte M,MaRty F, et al. Autophagy in development and stRess Responses of plants[J]. Autophagy, 2006,2 ( 1) : 2 - 11.
[4]  Bassham D C. Plant autophagy-moRe than a staRvation Response
[5]  [J]. CuRRent Opinion in Plant Biology,2007,10 ( 6) : 1 - 7
[6]  Yoshimoto K,JikumaRu Y,Kamiya Y,et al. Autophagy negatively Regulates cell death by contRolling NPR1-dependent salicylic acid signaling duRing senescence and the innate immune Response in ARabidopsis[J]. The Plant Cell, 2009, 21( 9) : 2914 - 2927.
[7]  Hofius D,Mundy J,PeteRsen M. Self-consuming innate immunity in ARabidopsis[J]. Autophagy,2009,5 ( 8 ) : 1206 - 1207.
[8]  HaywaRd A P,Tsao J,Dinesh-KumaR S P. Autophagy and plant innate immunity: Defense thRough degRadation[J]. SeminaRs in Cell & Developmental Biology, 2009, 20( 9) : 1041 - 1047.
[9]  Yue Z, Jin S,Yang C, et al. Beclin 1,an autophagy gene essential foR eaRly embRyonic development, is a haploinsufficient tumoR suppRessoR[J]. PRoceedings of the National Academy of Sciences, 2003, 100( 25) : 15077 - 15082
[10]  Liu W,Yan A H,Hou C Y, et al. Cloning and pRokaRyotic expRession of TaCaM-2-3 in wheat and pRepaRation of antiseRum[J]. FRontieRs of AgRicultuRe in China, 2010,4 ( 3) : 317 - 322.
[11]  Klionsky D J,CueRvo A M,Seglen P O. Methods foR monitoRing autophagy fRom yeast to human[J]. Autophagy, 2007,3 ( 3) : 181 - 206.
[12]  Mizushima N. Methods foR monitoRing autophagy[J]. Int J Biochem Cell Biol, 2004, 36( 12) : 2491 - 2502.
[13]  Klionsky D J,Abeliovich H,Agostinis P, et al. Guidelines foR the use and inteRpRetation of assays foR monitoRing autophagy in higheR eukaRyotes[J]. Autophagy, 2008,4 ( 2) : 151 - 175.
[14]  Klionsky D J. AutoPhagy: fRom Phenom enology to moleeulaR undeRstanding inless than a deeade[J]. Nat Revmol Cell Biol, 2007,8 ( 11) : 931 - 937.
[15]  Xie Z,Klinsky D J. Autophagsome fomRtion: coRe machineRy and adaptations[J]. Nat Cell Biol,2007,9 ( 10) : 1102 - 1109.
[16]  KiRisako T, IehimuRa Y,Okada H,et al. The ReveRsible modifieation Regulates the membRane-binding state of APg8 /Aut7 essential foR auto phagy and the cytoplasm to vacuole taRgeting Pathway[J]. J Cell Biol,2000,151 ( 2) : 263 - 276.
[17]  IchimuRa Y,KiRisako T,Takao T, et al. Aubiquitin-like systemmediates pRotein lipidation[J]. NatuRe, 2000, 408 ( 6811) : 488 - 492.
[18]  Xie Z,NaiR U,Klionsky D J. AtgseontRols phagophoRe expansion duRing autophagosome foRmation[J]. Mol Biol Cell, 2008, 19( 8) : 3290 - 3298.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133