Split-Cre Complementation Restores Combination Activity on Transgene Excision in Hair Roots of Transgenic Tobacco

	Split-Cre Complementation Restores Combination Activity on Transgene Excision in Hair Roots of Transgenic Tobacco
	Wen, Mengling 1,2; Gao, Yuan 1,2; Wang, Lijun 1,2; Ran, Lingyu 1,2; Li, Jiahui 1,2; Luo, Keming 1,2,3
刊名	plos one
	2014-10-17
卷号	9 期号:10
ISSN号	1932-6203
中文摘要	the cre/loxp system is increasingly exploited for genetic manipulation of dna in vitro and in vivo. it was previously reported that inactive "split-cre'' fragments could restore cre activity in transgenic mice when overlapping co-expression was controlled by two different promoters. in this study, we analyzed recombination activities of split-cre proteins, and found that no recombinase activity was detected in the in vitro recombination reaction in which only the n-terminal domain (ncre) of split-cre protein was expressed, whereas recombination activity was obtained when the c-terminal (ccre) or both ncre and ccre fragments were supplied. we have also determined the recombination efficiency of split-cre proteins which were co-expressed in hair roots of transgenic tobacco. no cre recombination event was observed in hair roots of transgenic tobacco when the ncre or ccre genes were expressed alone. in contrast, an efficient recombination event was found in transgenic hairy roots co-expressing both inactive split-cre genes. moreover, the restored recombination efficiency of split-cre proteins fused with the nuclear localization sequence (nls) was higher than that of intact cre in transgenic lines. thus, dna recombination mediated by split-cre proteins provides an alternative method for spatial and temporal regulation of gene expression in transgenic plants.
英文摘要	the cre/loxp system is increasingly exploited for genetic manipulation of dna in vitro and in vivo. it was previously reported that inactive "split-cre'' fragments could restore cre activity in transgenic mice when overlapping co-expression was controlled by two different promoters. in this study, we analyzed recombination activities of split-cre proteins, and found that no recombinase activity was detected in the in vitro recombination reaction in which only the n-terminal domain (ncre) of split-cre protein was expressed, whereas recombination activity was obtained when the c-terminal (ccre) or both ncre and ccre fragments were supplied. we have also determined the recombination efficiency of split-cre proteins which were co-expressed in hair roots of transgenic tobacco. no cre recombination event was observed in hair roots of transgenic tobacco when the ncre or ccre genes were expressed alone. in contrast, an efficient recombination event was found in transgenic hairy roots co-expressing both inactive split-cre genes. moreover, the restored recombination efficiency of split-cre proteins fused with the nuclear localization sequence (nls) was higher than that of intact cre in transgenic lines. thus, dna recombination mediated by split-cre proteins provides an alternative method for spatial and temporal regulation of gene expression in transgenic plants.
WOS标题词	science & technology
类目[WOS]	multidisciplinary sciences
研究领域[WOS]	science & technology - other topics
关键词[WOS]	site-specific recombination ; cre/loxp system ; alpha-complementation ; dna recombination ; selectable marker ; gene ; expression ; protein ; transformation ; sequences
收录类别	SCI
语种	英语
WOS记录号	WOS:000345204100057
公开日期	2014-12-19
内容类型	期刊论文
源URL	[http://ir.nwipb.ac.cn/handle/363003/4179]
专题	西北高原生物研究所_中国科学院西北高原生物研究所
作者单位	1.Southwest Univ, Key Lab Ecoenvironm Gorges Reservoir Reservoir Re, Sch Life Sci, Minist Educ,Inst Resources Bot, Chongqing, Peoples R China 2.Southwest Univ, Chongqing Key Lab Transgen Plant & Safety Control, Chongqing, Peoples R China 3.Chinese Acad Sci, Northwest Inst Plateau Biol, Key Lab Adaptat & Evolut Plateau Biota, Xining, Peoples R China
推荐引用方式 GB/T 7714	Wen, Mengling,Gao, Yuan,Wang, Lijun,et al. Split-Cre Complementation Restores Combination Activity on Transgene Excision in Hair Roots of Transgenic Tobacco[J]. plos one,2014,9(10).
APA	Wen, Mengling,Gao, Yuan,Wang, Lijun,Ran, Lingyu,Li, Jiahui,&Luo, Keming.(2014).Split-Cre Complementation Restores Combination Activity on Transgene Excision in Hair Roots of Transgenic Tobacco.plos one,9(10).
MLA	Wen, Mengling,et al."Split-Cre Complementation Restores Combination Activity on Transgene Excision in Hair Roots of Transgenic Tobacco".plos one 9.10(2014).