[1]侯永杰,李庆华,薛乐勋#.杜氏盐藻基因FLA8原核表达载体的构建和表达*[J].郑州大学学报(医学版),2013,(01):31.
 HOU Yongjie,LI Qinghua,XUE Lexun.Construction and expression of FLA8 from Dunaliella salina prokaryotic expression vector[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2013,(01):31.
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杜氏盐藻基因FLA8原核表达载体的构建和表达*
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2013年01期
页码:
31
栏目:
论著
出版日期:
2013-01-20

文章信息/Info

Title:
Construction and expression of FLA8 from Dunaliella salina prokaryotic expression vector
作者:
侯永杰李庆华薛乐勋#
郑州大学生物工程系细胞生物学研究室 郑州 450001
Author(s):
HOU YongjieLI QinghuaXUE Lexun
Laboratory for Cell Biology,Department of Bioengineering,Zhengzhou University,Zhengzhou 450001
关键词:
FLA8原核表达杜氏盐藻
Keywords:
FLA8prokaryotic expressionDunaliella salina
分类号:
Q786
摘要:
目的:构建杜氏盐藻FLA8原核表达载体。方法:用RTPCR方法扩增杜氏盐藻FLA8基因的开放阅读框,通过预先添加的酶切位点切割后按正确的顺序插入到经相同酶切割后的原核表达载体pET28a(+)中,转化到大肠杆菌DE3中测序正确后进行原核表达。结果:成功地将FLA8的开放阅读框插入到表达载体中,FLA8在大肠杆菌中的表达主要以包涵体的形式存在,蛋白质的相对分子质量为87 000。结论:杜氏盐藻FLA8基因原核表达载体构建成功。
Abstract:
Aim:To build recombination prokaryotic expression vector of FLA8 from Dunaliella salina.Methods:Open reading frame of the FLA8 was amplified by RTPCR,then inserted into prokaryotic expression vector pET28a(+) which was digested by the same enzyme and transformed into DE3 after sequencing.Results:The open reading frame of the FLA8 was inserted into the expression vector successfully and mainly in the form of inclusion bodies with relative molecular weight of 87 000.Conclusion:The prokaryotic expression vector of FLA8 from Dunaliella salina has been successfully constructed.

参考文献/References:

[1]Sawin KE,LeGuellec K,Philippe M,et al.Mitotic spindle organization by a plusenddirected microtubule motor[J].Nature,1992,359(6395):540
[2]Barton NR,Goldstein LS.Going mobile:microtubule motors and chromosome segregation[J].Proc Natl Acad Sci USA,1996,93(5):1735
[3]Moore JD,Endow SA.Kinesin proteins:a phylum of motors for microtubulebased motility[J]Bioessays,1996,18(3):207
[4]Wein H,Foss M,Brady B.DSK1,a novel kinesinrelated protein from the diaom Cylindrotheca fusiformis that is involved in anaphase spindle elongation[J].J Cell Biol,1996,133(3):595
[5]Raich WB,Moran AN,Rothman JH.Cytokinesis and midzone microtubule organization in Caenorhabditis elegans require the kinesinlike protein ZEM4[J].Mol Biol Cell,1998,9(8):2037
[6]李俊平,刘岷,毛丽红,等.杜氏盐藻FLA8基因全长的克隆及鉴定[J].郑州大学学报:医学版,2011,46(6):828
[7]Eley L,Yates LM,Goodship JA.Cilia and disease[J].Curr Opin Genet Develop,2005,15(3):308
[8]Marande W,Kohl L.Flagellar kinesin in protests[J].Future Microbiol,2011,6(2):231
[9]Pazour GJ,Witman GB.The vertebrate primary cilium is a sensory organelle[J].Curr Opin Cell Biol,2003,15(1):105
[10]Rosenbaum JL,Witman GB.Intraflagellar transport[J].Nat Rev Mol Cell Biol,2002,3(11):813
[11]Scholey JM.Intraflagellar transport[J].Annu Rev Cell Dev Biol,2003,19:423
[12]Qin H,Diener DR,Geimer S,et al.Intraflagellar transport(IFT)cargo:IFT transports flagellar precursors to the tip and turnover products to the cell body[J].J Cell Biol,2004,164(2):255

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备注/Memo

备注/Memo:
*国家自然科学基金资助项目30700014;科技部国际科技合作基金资助项目2007DFA01240 #通讯作者,男,1944 年2 月生,本科,教授,研究方向:肿瘤标志物与基因工程,Email:xuelx@zzu.edu.cn
更新日期/Last Update: 2013-04-19