Knapp S, Larondelle Y, Rossberg M, Furtek D, Theres K
Institut für Genetik, Universität zu Köln, Germany.
Mol Gen Genet. 1994 Jun 15;243(6):666-73. doi: 10.1007/BF00279576.
We have introduced a genetically marked Dissociation transposable element (DsHPT) into tomato (Lycopersicon esculentum) by Agrobacterium tumefaciens-mediated transformation. Probes for the flanking regions of the T-DNA and transposed DsHPT elements were obtained with the inverse polymerase chain reaction (IPCR) technique and used in RFLP linkage analyses. The RFLP map location of 11 T-DNAs carrying DsHPT was determined. The T-DNAs are distributed on 7 of the 12 tomato chromosomes. To explore the feasibility of gene tagging strategies in tomato using DsHPT, we examined the genomic distribution of DsHPT receptor sites relative to the location of two different, but very closely linked, T-DNA insertion sites. After crosses with plants expressing Ac transposase, the hygromycin phosphotransferase (HPT) marker on the Ds element and the excision markers beta-glucuronidase (GUS) and Basta resistance (BAR) facilitated the identification of plants bearing germinally transposed DsHPT elements. RFLP mapping of 21 transposed DsHPT elements originating from the two different T-DNA insertions revealed distinct patterns of reintegration sites.
我们通过根癌农杆菌介导的转化方法,将一个带有遗传标记的解离转座元件(DsHPT)导入番茄(Lycopersicon esculentum)。利用反向聚合酶链反应(IPCR)技术获得了T-DNA侧翼区域和转座后的DsHPT元件的探针,并将其用于RFLP连锁分析。确定了携带DsHPT的11个T-DNA的RFLP图谱位置。这些T-DNA分布在番茄12条染色体中的7条上。为了探索使用DsHPT在番茄中进行基因标签策略的可行性,我们研究了DsHPT受体位点相对于两个不同但紧密连锁的T-DNA插入位点位置的基因组分布。与表达Ac转座酶的植株杂交后,Ds元件上的潮霉素磷酸转移酶(HPT)标记以及切除标记β-葡萄糖醛酸酶(GUS)和巴斯塔抗性(BAR)有助于鉴定携带经生殖转座的DsHPT元件的植株。对源自两个不同T-DNA插入的21个转座后的DsHPT元件进行RFLP定位,揭示了重新整合位点的不同模式。