[1]许芳,石科,李靓,等.杜氏盐藻FKBP cDNA 的克隆及其在鞭毛解组装中的功能*[J].郑州大学学报(医学版),2011,(06):825.
 XU Fang,SHI Ke,LI Liang,et al.Cloning of FKBP cDNA from Dunaliella salina and analysis of its functions in flagellar disassembly[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2011,(06):825.
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杜氏盐藻FKBP cDNA 的克隆及其在鞭毛解组装中的功能*
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2011年06期
页码:
825
栏目:
专题研究
出版日期:
2011-11-20

文章信息/Info

Title:
Cloning of FKBP cDNA from Dunaliella salina and analysis of its functions in flagellar disassembly
作者:
许芳石科李靓张楠楠薛乐勋#
郑州大学生物工程系细胞生物学研究室 郑州 450002
Author(s):
XU FangSHI KeLI LiangZHANG NannanXUE Lexun
Laboratory for Cell Biology, Department of Bioengineering, Zhengzhou University,Zhengzhou 450002
关键词:
杜氏盐藻FK506结合蛋白鞭毛
Keywords:
Dunaliella salina FK506 binding proteinflagella
分类号:
Q781
摘要:
目的:克隆杜氏盐藻FK506结合蛋白(FKBP)的cDNA片段并探讨其与鞭毛微管解组装的联系。方法:提取杜氏盐藻总RNA,根据已知盐藻FKBP cDNA 3’端序列,用5’RACE方法扩增该cDNA的5’端序列,拼接得到全长并对其序列进行分析。秋水仙碱处理对数生长期杜氏盐藻,处理0、20、40、60、80、100、120、140和160 min后采用实时荧光定量PCR检测FKBP mRNA的表达情况。结果:经5’RACE得到681 bp的FKBP cDNA片段,拼接后得到的cDNA全长1 075 bp,序列分析显示其开放阅读框为762 bp,编码253个氨基酸;经BLAST比对,其氨基酸序列具有FKBP家族特有的功能结构域,与莱茵衣藻、团藻、小球藻、大麦及水稻的同源性分别为61%、54%、47%、48%和51%。杜氏盐藻细胞FKBP mRNA表达量在秋水仙碱处理后20~40 min明显升高,而在40~100 min时逐渐下降,但仍高于对照组(F组间=32.580,P<0.001;F时间=5.543,P=0.001;F交互=237.306,P<0.001)。结论:得到了杜氏盐藻的FKBP cDNA全长序列;FKBP参与了杜氏盐藻微管解组装过程,可能参与了细胞通过鞭毛对外界应激反应的应答。
Abstract:
Aim:To clone the cDNA fragment of FK506 binding protein(FKBP) from Dunaliella salina(D. salina) and to investigate its function in the process of microtubule disassembly in flagella.Methods:The total RNA was extracted from D.salina and the full length of FKBP cDNA was amplified by 5’RACE method according to the known 3 ’terminal sequence.The expression of FKBP mRNA in D. salina treated with colchicine for 0,20,40,60,80,100,120,140 and 160 min,was detected by realtime fluorescence quantitative PCR.Results:A 681 bp DNA fragment was obtained by 5’RACE. The full length of FKBP cDNA was 1 075 bp long and contained an open reading frame of 762 bp which encodes 253 amino acids. Sequence analysis showed that it had the conserved domain of FKBP family and was homologous with Chlamydomonas reinhardtii(61%),Volvox carteri f. nagariensis(54%),Chlorella variabilis(47%),Hordeum vulgare subsp.Vulgare(48%) and Oryza sativa Indica Group(51%). The expression of FKBP mRNA in D.salina 20~40 min after colchicine treatment significantly increased,and decreased gradually within 40~100 min after colchicine treatment,but still higher than the control (Fgroup=32.580,P<0.001;Ftime=5.543,P=0.001;Finteraction=237.306,P<0.001).Conclusion:The FKBP cDNA sequence of D.salina has been obtained successfully.The expression of FKBP has correlation with the process of microtubule disassembly in D. salina,and it may be involved in the response of the cells to external stress through the flagella.

参考文献/References:

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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,教授,博士研究生导师,研究方向:肿瘤标志物与基因工程,Email:xuelx@zzu.edu.cn
更新日期/Last Update: 1900-01-01