[1]唐蓓△.小鼠Arl8a基因逆转录病毒载体MSCVGFPArl8a的构建*[J].郑州大学学报(医学版),2009,(06):1204-1206.
 TANG Bei.Cloning of mouse Arl8a gene and construction of its retroviral vector[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2009,(06):1204-1206.
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小鼠Arl8a基因逆转录病毒载体MSCVGFPArl8a的构建*()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2009年06期
页码:
1204-1206
栏目:
论著
出版日期:
2009-11-01

文章信息/Info

Title:
Cloning of mouse Arl8a gene and construction of its retroviral vector
作者:
唐蓓△
重庆师范大学生命科学学院 重庆 400047
Author(s):
TANG Bei
College of Life Sciences,Chongqing Normal University, Chongqing 400047
关键词:
Arl8a克隆逆转录病毒载体小鼠
Keywords:
ADPribosylation factorlike 8Acloneretroviral vectormouse
分类号:
R392
摘要:
克隆小鼠Arl8a(mArl8a) cDNA,构建带有检测标签的逆转录病毒载体系统,并检测由该病毒系统介导的mArl8a基因在骨髓来源树突状细胞中的表达。方法:从培养的小鼠骨髓来源树突状细胞中提取总RNA,通过RTPCR扩增出mArl8a 基因片段,再将其克隆入pDrive克隆载体并测序。然后将mArl8a目的基因插入逆转录病毒载体MSCVGFP中,以磷酸钙法转染294T包装细胞,收集病毒上清感染树突状细胞后,通过流式细胞术检测mArl8a的表达。结果与结论:成功克隆了mArl8a的 cDNA,并成功构建其重组逆转录病毒载体MSCVGFPArl8a,该载体可有效将Arl8a基因转移到骨髓来源的树突状细胞中,转染效率达28.3%。
Abstract:
To clone mouse ADPribosylation factorlike 8A (mArl8a) gene,construct retroviral vectors,and explore the gene transfection efficiency of the vector in bone marrowderived dendritic cells (BMDCs).Methods:The mArl8a cDNA was amplified by RTPCR from total RNA derived from mouse BMDCs, and then was cloned into pDrive cloning vector. After being sequenced, the mArl8a was inserted into retroviral vector MSCVGFP to generate MSCVGFPArl8a.The recombinant vector was transfected into packaging cell 294T to generate the complete retrovirus which infect BMDCs. Expression of mArl8a in infected cells was analyzed by fluorescence activated cell sorter.Results and Conclusion: The mArl8a cDNA was cloned and MSCVGFPArl8a was constructed successfully. Recombinant vector could transfer effectively mArl8a genes into BMDCs, and the transfection efficiency was 28.3%.

参考文献/References:

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[3]Hofmann I, Munro S. An Nterminally acetylated Arflike GTPase is localized to lysosomes and affects their motility[J]. J Cell Sci,2006,119(Pt 8):1 494
[4]Bagshaw RD,Callahan JW,Mahuran DJ.The Arffamily protein,Arl8b,is involved in the spatial distribution of lysosomes[J].Biochem Biophys Res Commun,2006,344(4):1 186
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备注/Memo

备注/Memo:
*重庆市自然科学基金资助项目2008BB5069;重庆师范大学基金资助项目08×LB007 △女,1970年生,博士,教授,研究方向:分子生物学及免疫学,Email:xiaobt26@yahoo.com.cn
更新日期/Last Update: 2010-05-14