Max-Planck-Institut für Züchtungsforschung, D-5000 Köln 30, FRG.
EMBO J. 1984 Dec 1;3(12):2723-30. doi: 10.1002/j.1460-2075.1984.tb02202.x.
The two most abundant transcripts derived from TR-DNA within plant cells transformed by an octopine strain of Agrobacterium tumefaciens arise from divergent transcription, both originating within an 500 bp section of the T-DNA. Using a combination of subcloning and exonuclease digestion, a 479-bp DNA fragment, directly flanked by the initiation codons for the two adjacent open reading frames, was isolated. The resulting DNA fragment was fused, in both orientations, to the neomycin phosphotransferase (NPT II) gene of the transposon Tn5 prior to introduction into Nicotiana tabacum cells via the Ti plasmid. The intergenic fragment was found to initiate expression of the NPT II gene in either orientation as assayed by kanamycin resistance of the transformed plant tissue as well as by enzymatic assay of the NPT II gene product. The plasmids described here are potential selection-expression vectors for plant systems.
在被农杆菌属胭脂碱型菌株转化的植物细胞中，源自 TR-DNA 的两种最丰富的转录本源自不同的转录，两者均源自 T-DNA 的 500bp 区段内。通过亚克隆和核酸外切酶消化的组合，分离出一个直接由两个相邻开放阅读框的起始密码子侧翼的 479bp DNA 片段。将所得 DNA 片段以两种方向融合到转座子 Tn5 的新霉素磷酸转移酶（NPT II）基因之前，通过 Ti 质粒引入烟草细胞。通过转化植物组织的卡那霉素抗性以及 NPT II 基因产物的酶促测定，发现该基因间片段以两种方向均能启动 NPT II 基因的表达。此处描述的质粒是植物系统的潜在选择表达载体。
