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piggyBac共转染系统的构建以及转座活性的验证 被引量：1

Constructing of a piggyBac binary cotransfection system and verifying the activity
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摘要 piggyBac(PB)转座子的末端反向重复序列(ITR)和转座酶编码序列(PBase)分别克隆于质粒PB1156、atub-pBac-K10.1TR与包含红色荧光蛋白表达序列(RFP)的质粒pmClherry-N1重组构成供体质粒(Donor);PBase与真核表达载体pEFrF-PGK重组构成辅助质粒(Helper).使用由供体质粒与辅助质粒组成的PB二元共转染系统转染HEK293细胞,结果表明该二元共转染系统具有较好的转座活性,并且在分子水平验证了转座的发生以及使用反向PCR(IPCR)定位了其中一个转座插入位点在HEK293基因组DNA上的位置. The PB inverted terminal repeats （ITR） and PB transposase （PBase） were cloned from the plasmid PBl156 and atub-pBac-K10. The PB binary cotransfection system consisted of a donor and a helper plasmid. The donor plasmid contained the two ITRs in which PBase was replaced by a red fluorescent protein （RFP） marker. The helper plasmid carried the PBase fragment but lacked the ITRs. The result of cotransfection the donor and helper in HEK293 cells indicated that the PB binary cotransfection system is an efficient transposition system. And inverse PCR （IPCR） was performed to recover sequence adjacent to the PB ITR.
作者 缪辉 刘华华 吴传芳
机构地区 四川大学生命科学学院功能基因组实验室
出处 《四川大学学报（自然科学版）》 CAS CSCD 北大核心 2011年第4期943-948,共6页 Journal of Sichuan University(Natural Science Edition)
基金 国家自然科学基金重大研究计划(90919005) 教育部博士点基金(20090181120076)
关键词 转座子 piggyBac(PB) 共转染 IPCR Transposon, piggyBac（PB）, Cotransfection,IPCR
分类号 Q789 [生物学—分子生物学]
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参考文献16

  • 1Lander E S, Linton L M, Birren B, et al. Initial se- quencing and analysis of the human genome [J]. Na- ture. 2001; 409: 860.
  • 2McClintock B. The origin and behavior of mutable lo- ci in Maize [J]. Proc Natl Aead Sci U S A, 1950, 36 : 344.
  • 3Hutchison C A, Peterson S N, Gill S R, etal. Glob- al transposon mutagenesis and a minimal mycoplasma genome [J]. Science, 1999, 286: 2165.
  • 4Rubin G M, Spradling A C. Genetic transformation of Drosophila with transposable element vectors [J]. Science, 1982, 218: 348.
  • 5Fraser M J, Hink W F. The isolation and character- ization of MP and FP plaque variants of galleria mel- lonella nuclear polyhedrosis virus [J]. Virology, 1982, 117:366.
  • 6Fraser M J, Smith G E, Summers M D. The acquisi- tion of host cell DNA sequences by Baculoviruses: Relation between host DNA insertions and FP mu- tants of Autographa californica and Galleria mellonel- la NPVs [J]. Virology, 1983, 47: 287.
  • 7Ding S, Wu X, Li G, et al. Efficient transposition of the piggyBac (PB) transposon in mammalian cells and mice [J]. Cell, 2005, 122: 473.
  • 8Herron B J, Lu W, Rao C, et al. Efficient genera- tion and mapping of recessive developmental muta-tions using ENU mutagenesis [J]. Nature Genetics, 2002, 30: 185.
  • 9Dupuy A J, Fritz S, Largaespada D A. Transposition and gene disruption in the male germline of the mouse [J]. Genesis, 2001, 30: 82.
  • 10Cooley L, Kelley R, Spradling A. Insertional muta- genesis of the Drosophila genome with single P ele- ments [J]. Science, 1988, 239: 1121.

二级参考文献1

共引文献4

同被引文献18

  • 1Ding S, Wu X, Li G, et al. Efficient transposition of the piggyBac (PB) transposon in mammalian cells and mice I-J3. Cell, 2005, 122(3), 473.. 483.
  • 2Wang W, Lin C, Lu D, et al. Chromosomal trans- position of piggyBac in mouse embryonic stem cells [J]. Proc Natl Acad Sci USA, 2008, 105 (27), 9290: 9295.
  • 3Rad R, Rad L, Wang W, etal. PiggyBac Transpo- son Mutagenesis: A Tool for Cancer Gene Discov- ery in Mice I-J, Science, 2010, 330 (6007), 1104: 1107.
  • 4Meir Y J, Weirauch T W, Yang H S, et al. Ge- nome-wide target profiling of piggyBac and To12 in HEK 293: pros and cons for gene discovery and gene therapy [J]. BMC Biotechnology, 2011, 11: 28.
  • 5Liang Q, Kong J, Stalker J, et al. Chromosomal mobilization and reintegration of Sleeping Beauty and PiggyBac transposons [J]. Genesis, 2009, 47 (6), 404: 408.
  • 6Li M A, Pettitt S J, Eckert S, et al. The piggybac transposon displays local and distant reintegration preferences and can cause mutations at non-canonical integration sites [J]. Mol Cell Biol, 2013, 33(7), 1317: 1330.
  • 7Ley D, Van Zwieten R, Puttini S, et al. A Piggy- Bac-mediated approach for muscle gene transfer or cell therapy [J]. Stem Cell Research, 2014, 13(3)390: 403.
  • 8Matsui H, Fujimoto N, Sasakawa N, et al. Deliv- ery of full-length factor " using a piggyback tra-ns- poson vector to correct a mouse model of hemophilia A[J]. PLoSOne, 2014, 9(8), e104957.
  • 9Wilson M H, Coates C J, George A L Jr. PiggyBac transposon-mediated gene transfer in human cells [J]. Molecular Therapy, 2007, 15(1), 139145.
  • 10Di Matteo M, Samara-Kuko E, Ward N J, et al. Hyperactive PiggyBac Transposons for Sustained and Robust Liver-targeted Gene Therapy[J]. Mo- lecular Therapy, 2014, 22(9), 1614: 1624.

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四川大学学报（自然科学版）
2011年 第4期

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