酵母双杂交技术筛选与STING蛋白相互作用的蛋白Screening for proteins interacting with protein kinase STING by yeast two hybrid system
刘晶莹;李胜龙;宋佳;富丹;
摘要(Abstract)：
目的应用酵母双杂交技术筛选出与干扰素刺激因子(stimulator of IFN genes,STING)相作用的分子,为深入研究STING蛋白的分子水平的功能奠定基础。方法利用基因克隆的方法构建酵母双杂交质粒pGBKT7-STING。应用酵母杂交技术从人胚肾cDNA库中筛选出与STING蛋白相互作用的蛋白,并在酵母中从新验证其相互作用。结果应用酵母双杂交系统筛选出与STING蛋白相作用的19个候选分子,成功构建pGBKT7-STING质粒,且可以在酵母中表达。结论应用酵母双杂交技术可筛选出与STING蛋白相互作用蛋白。
关键词(KeyWords)： STING;酵母双杂交;蛋白互作
基金项目(Foundation): 牡丹江市科学技术计划项目(Z2015s0025)
作者(Authors): 刘晶莹;李胜龙;宋佳;富丹;
DOI: 10.13799/j.cnki.mdjyxyxb.2019.03.008
参考文献(References)：
- [1] ZHANG Y,LIANG C.Innate recognition of microbial-derived signals in immunity and inflammation,science china[J].Sci China Life Sci,2016,59(12):1210-1217.
- [2] WU J,CHEN Z J.Innate immune sensing and signaling of cytosolic nucleic acids[J].Annu,2017,32(8):652-675.
- [3] ISHIKAWA H,BARBER G N.STING is an endoplasmic reticulum adaptor that facilitates innate immune signaling[J].Nature,2009,48(1/2):128-128.
- [4] SAUER J D,SOTELOT R K,VON M J,et al.The N-ethyl-N-nitrosourea-induced Golden ticket mouse mutant reveals an essential function of Sting in the in vivo interferon response to Listeria mono cyto genes and cyclic dinucleotides[J].Infect Immun,2011,79(2):688-694.
- [5] BURDETTE D L,MONROE K M,SOTELOTROH A K,et al.STING is a direct innate immune sensor of cyclic-di -GMP[J].Nature,2011,478(7370):515-518.
- [6] CONLON J,BURDETTE D L,SHARMA S,et al.Mouse,but not human STING,binds and signals in response to the vascular disrup-ting agent 5,6-dimethylxan the none- 4-acetic acid[J].J Immunol,2013,190(10):5216-5225.
- [7] ZHONG B,YANG Y,LI S,et al.The a daptor protein MITA links virus-sensing receptors to IRF3tran-scription factor activation[J].Immunity,2008(29):538-550.
- [8] TANER C,TOBIAS D,ABLASSER A,et al.Species-specific detection of the antiviral small-molecule compound CMA by STING[J].EMBO J,2013,32(10):1440-1450.
- [9] CIVRIL T M,DEIMLING T O,RAMANN C C,et al.Struc-tural mechanism of cytosolic DNA sensing by c GAS[J].Nature,2013,498(7454):332-337.
- [10] TSUCHIYA Y,JOUNAI N,TAKESHITA F,et al.Ligand-induced ordering of the c-terminal tail primes STING for phosphorylation by TBK1[J].EBio Medicine,2016(9):87-96.
- [11] MUELLER S,HAAS-KOGAN D A.WEE1 Kinase As a Target for Cancer Therapy[J].J Clin Oncol,2015,33(30):3485-3487.
- [12] GUZMáN V M,RINCON S A,DINGLI F,et al.Molecular control of the STING regulatory pathway by the SAD kinase Cdr2[J].J Cell Sci,2015,128(15):2842-2853.