期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

改进重叠延伸PCR克隆tau基因6种同工异构体 被引量：3

Cloning of Six Isoforms of Tau by Modified Overlap Extension PCR Technique
在线阅读 下载PDF
导出
摘要 采用一种改进的重叠延伸PCR技术,通过2次PCR的方法,对人tau基因进行剪切和拼接,实现了tau基因的6种同工异构体的克隆;然后将6种tau基因克隆到真核载体pcDNA4上,通过western blot在蛋白水平验证了tau蛋白的表达.结果表明,这种改进重叠延伸PCR方法能够对基因进行多个位置的剪切和连接,由于该法在基因重组过程中无需利用限制性内切酶,因此具有很大的灵活性和简便性. Here we describe a new modified overlap extension PCR method for cloning human tau isoforms which can cut and splice gene to make inserts for expression vectors easily.The isoforms could be simply cloned to eukaryotic expression vector pcDNA4 and the tau protein expression be detected by western blot.Our data implicate that this method is much convenience and easier to cut and splice the multiple cloning site without restriction enzyme.It possesses great flexibility,valuablity and may provide broad applications in molecular cloning.
作者 李诺敏 刘可夫 李灵芸 何放 顾文彬 董一名 邓玉林 庆宏
机构地区 北京理工大学生物医药成分分离与分析北京市重点实验室
出处 《北京理工大学学报》 EI CAS CSCD 北大核心 2013年第8期871-875,共5页 Transactions of Beijing Institute of Technology
基金 国家自然科学基金资助项目(81171206) 大学生创新性实验计划资助项目(BJ1160)
关键词 TAU蛋白 重叠延伸PCR 真核表达载体 TA克隆 tau protein overlap extension PCR eukaryotic expression vector TA cloning
分类号 Q812 [生物学—生物工程]
  • 相关文献

参考文献12

  • 1Weingarten M D, Lockwood A H, Hwo S Y, et al. A protein factor essential for microtubule assembly [J]. Proc Natl Acad Sci USA, 1975,72(5):1858- 1862.
  • 2Drechsel D N, Hyman A A, Cobb M H, et al. Modulation of the dynamic instability of tubulin assembly by the microtubule-associated protein tau [J]. Mol Biol Cell, 1992,3(10) :1141 - 1154.
  • 3Grundke-Iqbal I, Iqbal K, Quinlan M, et al. Microtubule-associated protein tau. a component of Alzheimer paired helical filaments [ J ]. J Biol Chem, 1986,261 (13) : 6084 - 6089.
  • 4Grundke-Iqbal I, Iqbal K, Tung Y C, et al. Abnormal phosphorylation of the microtubule-associated protein tau (tau) in Alzheimer cytoskeletal pathology [J]. Proc Natl Acad Sci U S A, 1986, 83(13): 4913-4917.
  • 5Goedert M, Spillantini M G, Jakes R, et al. Multiple isoforms of human microtubule-associated protein tau: sequences and localization in neurofibrillary tangles of Alzheimer's disease[J]. Neuron, 1989,3(4) :519 - 526.
  • 6Andreadis A, Brown W M, Kosik K S. Structure and novel exons of the human-tau gene [J]. Biochemistry, 1992,31 (43) : 10626 - 10633.
  • 7Kosik K S, Orecchio L D, Bakalis S, et al. Developmentally regulated expression of specific tau sequences[J]. Neuron, 1989,2(4) :1389 - 1397.
  • 8Goedert M, Jakes R. Expression of separate isoforms of human tau protein: correlation with the tau pattern in brain and effects on tubulin polymerization[J]. EMBO J, 1990,9(13): 4225-4230.
  • 9Goedert M, Spillantini M G, Potier M C, et al. Cloning and sequencing of the cDNA encoding an isoform of mi- crotubule-associated protein tau containing four tandem repeats: differential expression of tau protein mRNAs in human brain [J]. EMBO J, 1989,8(2):393 - 399.
  • 10Himmler A, Drechsel D, Kirschner M W, et al. Tau consists of a set of proteins with repeated C-terminal microtubule-binding domains and variable N-terminal domains[J]. Molecular and cellular biology, 1989, 9(4) :1381 - 1388.

同被引文献25

  • 1Kobayashi T, Nishizawa NIL Iron uptake, translocation and regulation in higher plants [J]. Annual Review ofPlan B/ogogy,2012,63:131 - 152.
  • 2Briat J F,Dubos C,Gaymard F. Iron nutrifion,biomass production,and plant product quality [J]. Trends in Plant Science, 2014, doi: http:// dx. doi. org/10. 1016/j. tplants. 2014. 07. 005.
  • 3HalliweU B, Gutteridge J M C. Biologically relevant metal ion - de- pendent hydroxyl radical generation an update [J]. FEBS Letters, 1992, 307 : 108 - 112.
  • 4La Fontaine S,Quinn J M,Nakamoto S S,et al. Copper- dependent i- ron assimilation pathway in the model photosynthetic eukaryote Chlamydo- monas reinhardtii [ J]. Eukaryoti Ce11,2002, 1 ( 5 ) :736 - 757.
  • 5Terzilli A, Kasman D J. Analysis of the high - affinity iron uptake sys- tem at the Chlamydomonas reinhardtii plasma membrane [ J]. Eukaryotic CeU,2010,9(5 ) :815 - 826.
  • 6Harris E H. The Chlamydomonas sourcebook [ M]. San Diego:Aca- demic Press, 1989:780.
  • 7Siflow C D. Organization of the nuclear genome [D]. In:Rochaix J D, Goldschmiddt - Clermont M, Merchant S ( eds ). The Molecular biology of chloroplast and mitochondria in Chlamydomonas [ M]. The Netherlands ( Dordrecht ) : Kluer Academic Publishers, 1998:25 - 40.
  • 8Garcia- Mina J, Bacaicoa E, Fuentesa M, Casanovaa E. Fine regula- tion of leaf iron use efficiency and iron root uptake under limited iron bio- availability [J ]. Plant Science,2013,198:39 - 45.
  • 9Singh B B,Patel H H, Roepman R, et al. The zinc finger cluster do- main of RanBP2 is a specific docking site for the nuclear export factorexpor- tin - 1 [ J]. The Journal of Biological Chemistry, 1999,274 ( 52 ) : 37370 - 37378.
  • 10Yaseen N R, Blobel G. Two distinct classes of Ran - binding sites on the nucleoporin Nup -358 [J]. Proc Natl Acad Sci USA,1999,96(10) : 5516 -5521.

引证文献3

二级引证文献5

北京理工大学学报
2013年 第8期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部