水牛Keap1基因的克隆、序列分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2016.08.007

Abstract

Abstract:

In this study,the CDS sequence of buffalo Keap1 gene was cloned and analyzed,then its expression pattern in different tissues was also investigated.A pair of primers of buffalo Keap1 gene was designed based on the nucleotide sequence of Bos taurus Keap1 gene from GenBank,and then the buffalo Keap1 gene was amplified.Using the bioinformation techniques,the gene sequence and the protein structure were analyzed.The expression level of Keap1 gene in different tissues were detected with Real-time quantitative PCR.The results showed that the length of buffalo Keap1 gene coding sequence was 1 875 bp and encoded 624 amino acids.The multiple sequence alignment results showed that buffalo Keap1 gene shared 99%,96%,92% and 90% of similar nucleotide sequence with that of Bos taurus,Ovis aries,Sus scrofa and Homo sapiens,respectively.And the phyogenetic tree also showed the conservatism between several different species.The second structure of buffalo Keap1 protein was predicted as 24 alpha regions,40 beta regions,38 turn regions and 27 coil regions.In addition,the results of Real-time quantitative PCR showed that Keap1 mRNA exists in all seven tissues,but the most abundant expression was in heart and the minimal expression was in liver and spleen.The results provided an foundation for further study of Keap1-Nrf2-ARE signal pathway,for enhancing the ability of antioxidant of buffalo embryo in vitro culture.

Key words: buffalo; Keap1 gene; clone; analysis; expression

CLC Number:

ZHAO Xin, RUAN Zi-yun, GUO Zhen-wei, ZHOU Wen-ting, QIN Xi-ling, NIU Xiang-li, LU Feng-hua, SHI De-shun. Cloning and Sequence Analysis of Buffalo Keap1 Gene and Investigation of Its Expression Pattern in Different Tissues[J]. , 2016, 43(8): 1975-1982.

References

[1] Kobayashi M,Itoh K,Suzuki T,et al.Identification of the interactive interface and phylogenic conservation of the Nrf2-Keap1 system[J].Genes Cells,2002,7(8):807-820.
[2] Itoh K,Wakabayashi N,Katoh Y,et al.Keap1 represses nuclear activation of antioxidant responsive elements by Nrf2 through binding to the amino-terminal Neh2 domain[J].Genes Dev Gene Dev,1999,13(1):76-86.
[3] Donna D Z.Mechanistic studies of the Nrf2-Keap1 signaling pathway[J].Drug Metab Rev,2006,38(4):769-789.
[4] 童海达,王佳茗,宋英.Keap1-Nrf2-ARE在机体氧化应激损伤中的防御作用[J].癌变·畸变·突变,2013,1:71-75.
[5] Wakabayashi N,Itoh K,Takahashi S.Keap1 mutation leads to postnatal lethality due to constitutive Nrf2 activation[J].Nat Genet,2003,35(3):238-245.
[6] 梁明振,杨炳壮,苏安伟,等.水牛奶营养价值评价[J].广西畜牧兽医,2007,3:124-126.
[7] 肖开田,张世生,李跃民.水牛繁殖生物技术研究进展[J].动物医学进展,2005,3:10-12.
[8] 庄利利,胡德宝,蔡丽娟,等.哺乳动物胚胎的活性氧来源及其抗氧化机制[J].中国畜牧兽医,2013,40(3):173-176.
[9] Dinkova-Kostova A T,Holtzclaw W D,Cole R N,et al.Direct evidence that sulfhydryl groups of Keap1 are the sensors regulating induction of phase 2 enzymes that protect against carcinogens and oxidants[J].P Natl Acad Sci USA,2002,99(18):11908-11913.
[10] 刘芳,杜志银,王应雄.Keap1在氧化应激方面的研究进展[J].中国临床药理学与治疗学,2010,5:596-600.
[11] Kansanen E,Jyrkkänen H K,Levonen A L.Activation of stress signaling pathways by electrophilic oxidized and nitrated lipids[J].Free Radical Biology & Medicine,2012,52(6):973-982.
[12] Kensler T,Wakabayashi N S.Cell survival responses to environmental stresses via the Keap1-Nrf2-ARE pathway[J].Annual Review of Pharmacology,2007,47:89-116.
[13] Hideaki K,Yoshihisa N,Takeshi M,et al.PDX-1/VP16 fusion protein,together with NeuroD or Ngn3,markedly induces insulin gene transcription and ameliorates glucose tolerance[J].Diabetes,2005,54(4):1009-1022.
[14] Kwak J J,Tirumani S H,Van den Abbeele A D,et al.Cancer immunotherapy:Imaging assessment of novel treatment response patterns and immune-related adverse events[J].Radiographics,2015,35(2):424-437.
[15] Xu C,Huang M G,Yuan X,et al.Inhibition of 7,12-dimethylbenz(a) anthracene-induced skin tumorigenesis in C57BL/6 mice by sulforaphane is mediated by nuclear factor E₂-related factor 2[J].Cancer Res,2006,66(16):8293-8296.
[16] Copple I M.The Keap1-Nrf2 cell defense pathway——A promising therapeutic target[J].Advances in Pharmacology,2012,63:43-79.
[17] Ishikawa M,Numazawa S,Yoshida T.Redox regulation of the transcriptional repressor Bach1[J].Free Radical Bio Med,2005,38(10):1344-1352.
[18] Maher J,Yamamoto M.The rise of antioxidant signaling——The evolution and hormetic actions of Nrf2[J].Toxicol Appl Pharm,2010,244(1):4-15.
[19] Zhang D D,Lo S C,Cross J V,et al.Keap1 is a redox-regulated substrate adaptor protein for a Cul3-dependent ubiquitin ligase complex[J].Mol Cell Biol,2004,24(24):10941-10953.
[20] Shibata T,Ohta T,Tong K I,et al.Cancer related mutations in NRF2 impairits recognition by Keap1-Cul3 E₃ ligaseand promote malignancy[J].PNAS,2008,105(36):13568-13573.
[21] Kim S K,Yang J W,Kim M R,et al.Increased expression of Nrf2/ARE-dependent anti-oxidant proteins in tamoxifen-resistant breast cancer cells[J].Free Radical Bio Med,2008,45(4):537-546.

Cloning and Sequence Analysis of Buffalo Keap1 Gene and Investigation of Its Expression Pattern in Different Tissues

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