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表达人WDR70基因的质粒构建及鉴定

Construction and Identification of Hunman WDR70-expressing Plasmid
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摘要 WD重复蛋白(WD repeat protein)家族是一类含有多个保守WD基序的蛋白.该家族蛋白广泛存在于真核细胞中,参与细胞发育、增殖和凋亡等生理过程.WDR70隶属于WD重复蛋白家族,其生物学功能目前仍不清楚.本文以人cDNA为模板扩增WDR70基因;然后以pTag2b为载体,通过同源重组构建重组质粒并进行测序.同时将质粒转染至人肺癌细胞株A549中,进行WDR70蛋白的表达分析.结果显示该重组质粒在A549细胞中能有效表达.质粒pTag2B-WDR70的成功构建为进一步研究WDR70相关生物学功能提供实验基础. WD repeat protein is a class of proteins containing several conservative WD motif. The family of WD protein extensively exists in eukaryocytes and involved in a variety of biological process,such as cell development,proliferation and apoptosis. WDR70 belongs to WD repeat protein family. At present,the function of WDR70 is not clear. In this experiment,PCR was performed on amplification of WDR70 gene. Then we constructed p CMV-Tag2B-WDR70 recombinant plasmid by homologous recombination. The recombinant plasmid was sequenced and transfected into A549 human lung cancer cells. Finally Western blot was performed to analysis the expression of WDR70 protein. The experimental results showed that WDR70 gene was successfully cloned,and p CMV-Tag2BWDR70 recombinant plasmid was expressed efficiently in A549 cells. The construction of p CMV-Tag2B-WDR70 recombinant plasmid provides experiment materials for further reserch on WDR70-related biological functions.
出处 《赣南师范学院学报》 2016年第3期58-60,共3页 Journal of Gannan Teachers' College(Social Science(2))
基金 国家自然科学基金青年基金项目(81402304) 赣南师范学院招标课题(15zb08)
关键词 WDR70 WD重复蛋白 CDNA克隆 WDR70 WD repeat protein cDNA cloning
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参考文献13

  • 1Smith TF,Gaitatzes C,Saxena K,et al.The WD repeat:a common architecture for diverse functions[J].Trends Biochem Sci,1999,24(5):181-185.
  • 2Li D,Roberts R.WD-repeat proteins:structure characteristics,biological function,and their involvement in human diseases[J].Cell Mol Life Sci,2001,58(14):2085-2097.
  • 3Smith TF.Diversity of WD-repeat proteins[J].Subcell Biochem,2008,48:20-30.
  • 4Volk A,Crispino JD.The role of the chromatin assembly complex(CAF-1)and its p60 subunit(CHAF1b)in homeostasis and disease[J].Biochim Biophys Acta,2015,1849(8):979-986.
  • 5Grimmel M,Backhaus C,Proikas-Cezanne T.WIPI-Mediated Autophagy and Longevity[J].Cells,2015,4(2):202-217.
  • 6Mahmoudi S,Henriksson S,Farnebo L,et al.WRAP53 promotes cancer cell survival and is a potential target for cancer therapy[J].Cell Death Dis,2011,2:e114.
  • 7Sato N,Koinuma J,Fujita M,Hosokawa M,et al.Activation of WD repeat and high-mobility group box DNA binding protein 1 in pulmonary and esophageal carcinogenesis[J].Clin Cancer Res,2010,16(1):226-239.
  • 8Sandrini F,Farmakidis C,Kirschner LS,et al.Spectrum of mutations of the AAAS gene in Allgrove syndrome:lack of mutations in six kindreds with isolated resistance to corticotropin[J].J Clin Endocrinol Metab,2001,86(11):5433-5437.
  • 9单宏爽,郭丽丽,慈宏亮,李亦平.人类新基因WDR70的克隆及初步研究[J].微生物学杂志,2005,25(1):11-15. 被引量:4
  • 10Wang L,Ye X,Liu Y,et al.Aberrant regulation of FBW7 in cancer[J].Oncotarget,2014,5(8):2000-2015.

二级参考文献7

  • 1Wilkinson D G, In situ hybridization. In: Stern CD, Holland PWH(eds.) Essential Development Biology: A Pratical Approach[M]. IRL Press, Oxford, UK, 1993,257-274.
  • 2Fong H. K., Hurley J. B., Hopkins R. S.,et al.Repetitive segmental structure of the transducin beta subunit: homology with the CDC4 gene and identification of related mRNAs[J].Proc. Natl. Acad. Sci. USA,1986, 83:2 162-2 166.
  • 3Yu L., Gaitatzes C., Neer E., et al. Thirty-plus functional families from a single motif[J]. Protein Sci, 2000,9:2 470-2 476.
  • 4Smith T. F. , Gaitatzes C., Saxena K. and Neer E. J. The WD repeat: a common architecture for diverse functions[J]. Trends Biochem. Sci, 1999,24:181 - 185.
  • 5D. Li and R. Robert. WD-repeat proteins:structure characteristics, biological function, and their involvement in human diseases[J]. Cell. Mol. Life Sci,2001,58:2 085 - 2 097.
  • 6Kawai J, Shinagawa A, Shibata K, Yoshino M, et al. Functional annotation of a full-length mouse cDNA collection [J]. Nature,2001,409 ( 6 821 ): 685 - 690.
  • 7李铁石,常智杰,傅新元,刘力.基于Internet网生物信息资源特定基因同源新基因克隆策略[J].生命科学,2002,14(4):234-237. 被引量:6

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