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Heat-Shock-Induced Removal of Transgenes Using the Gene-Deletor System in Hybrid Aspen (Populus tremula x P. tremuloides)

文献类型： 外文期刊

作者： Wang, Beibei 1 ; Zhang, Yan 1 ; Zhao, Jian 1 ; Dong, Mingliang 1 ; Zhang, Jinfeng 1 ;

作者机构： 1.Beijing Forestry Univ, Coll Biol Sci & Technol,Minist Educ,Key Lab Fores, Beijing Adv Innovat Ctr Tree Breeding Mol Design, Natl Engn Lab Tree Breeding,Key Lab Genet & Breed, Beijing 100083, Peoples R China

2.Beijing Acad Forestry & Pomol Sci, Beijing 100093, Peoples R China

关键词： gene-deletor system; Populus; heat shock; methylation; expression

期刊名称：GENES （ 影响因子：4.096； 五年影响因子：4.339 ）

ISSN：

年卷期： 2018 年 9 卷 10 期

页码：

收录情况： SCI

摘要： To evaluate the efficacy of the gene-deletor system in aspen, we evaluated the system for foreign gene removal in a hybrid aspen clone, INRA 353-53 (Populus tremula x P. tremuloides). The recombinase flipping DNA (FLP) gene was under the control of the heat-inducible promoter of Gmhsp17.6-L, and the beta-glucuronidase (gusA) gene which was under the control of the 35S promoter and were constructed using the gene-deletor system in the pCaLFGmFNLFG vector. Six transgenic plants and their sublines were heated at 42 degrees C for 8 h and gene deletion was verified by polymerase chain reaction (PCR). Three lines exhibited partial transgene deletion while the remaining three lines did not delete. Transgenic lines were evaluated by Southern-blot analyses, verifying that the six transgenic plant lines all had a single copy of transfer DNA (t-DNA). Two partial-deletion lines and two non-deletion lines were analysed for methylation and expression of promoter and recombinase. Hardly any methylation was detected in the Gmhsp17.6-L promoter or recombinase FLP gene sequences, however, the expression of the promoter and recombinase was increased significantly in the partial-deletion compared with the non-deletion line after heat-shock treatment. These results suggest that the excision efficiency had no direct relationship with methylation status of the Gmhsp17.6-L promoter and FLP recombinase, yet was affected by the expression of the Gmhsp17.6-L and FLP after heat-shock treatment.

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