EFFICIENT OCTOPINE TI PLASMID-DERIVED VECTORS FOR AGROBACTERIUM-MEDIATED GENE-TRANSFER TO PLANTS
EFFICIENT OCTOPINE TI PLASMID-DERIVED VECTORS FOR AGROBACTERIUM-MEDIATED GENE-TRANSFER TO PLANTS
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DOI：
10.1093/nar/13.13.4777
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发表时间：
1985-01-01
影响因子：
14.9
通讯作者：
LEEMANS, J
中科院分区：
文献类型：
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作者：
DEBLAERE, R;BYTEBIER, B;LEEMANS, J
以章鱼碱Ti质粒pTiB 6S 3为载体，建立了一个双组分克隆系统。pGV 2260是一种非致癌Ti质粒，其中T区缺失并被pBR 322取代。pGVB 31是链霉素抗性pBR 325衍生物，其含有植物细胞的卡那霉素抗性标记基因和在章鱼碱T区的25-bp边界序列之间克隆外源基因的位点。pGV 831衍生物与农杆菌的接合转移和pGV 2260与pGV 831上的pBR 322序列之间的同源重组共整合可以在一个步骤中获得。携带所得共整合质粒的菌株将T-DNA转移并整合到烟草原生质体的基因组中，并且转化的烟草愈伤组织容易被选择为对卡那霉素具有抗性。从这样的克隆中已经再生出在T-DNA边界之间含有整个DNA区域的完整植物。鉴于这些性质，我们提出pGV 831及其衍生物作为载体，有效地整合外源基因到植物中。
A two-component cloning system to transfer foreign DNA into plants was derived from the octopine Ti plasmid pTiB6S3. pGV2260 is a non-oncogenic Ti plasmid from which the T-region is deleted and substituted by pBR322. pGVB31 is a streptomycin-resistant pBR325 derivative that contains a kanamycin resistance marker gene for plant cells and a site for cloning foreign genes between the 25-bp border sequences of the octopine T-region. Conjugative transfer of pGV831 derivatives toAgrobacteriumand cointegration by homologous recombination between the pBR322 sequences present on pGV831 and pGV2260, can be obtained in a single step. Strains carrying the resulting cointegrated plasmids transfer and integrate T-DNA into the genome of tobacco protoplasts, and transformed tobacco calli are readily selected as resistant to kanamycin. Intact plants containing the entire DNA region between the T-DNA borders have been regenerated from such clones. In view of these properties we present pGV831 and its derivatives as vectors for efficient integration of foreign genes into plants.