A novel Ti-plasmid-convertible lambda phage vector system suitable for gene isolation by genetic complementation of Arabidopsis thaliana mutants.
A new lambda phage vector system, lambda TI, has been constructed to facilitate genetic complementation of higher plant mutations. The lambda TI vectors are stable, and by using the Cre-lox site-specific recombination, are automatically convertible into Ti-plasmid binary vectors which are capable of expressing genes in higher plants. Two lambda TI vectors were constructed: (i) lambda TI1, which can generate a Ti-plasmid that contains the cauliflower mosaic virus (CaMV) 35S promoter and is suitable for the expression of cDNA in transformed plants and (ii) lambda TI2, which can generate a Ti-plasmid with the multicloning site (MCS). cDNA and genomic libraries, which were constructed from the cruciferous plant Arabidopsis thaliana in these lambda TI vectors, can be probed by large DNA fragments of more than 100 kb, such as yeast artificial chromosomes (YACs), enabling the direct screening of the clones in the chromosome region containing a specified genetic locus. These libraries will certainly become powerful tools for the genetic complementation of Arabidopsis mutant phenotypes by quickly providing transformation-competent clones.
Fuse
T
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Department of Biology, Faculty of Science, Kyushu University, Fukuoka, Japan.
Kodama
H
H
Hayashida
N
N
Shinozaki
K
K
Nishimura
M
M
Iba
K
K
D016960:Agrobacterium tumefaciens / Q000235:genetics; D017360:Arabidopsis / Q000235:genetics*; D010582:Bacteriophage lambda / Q000235:genetics*; D001483:Base Sequence; D017796:Caulimovirus / Q000235:genetics; D018244:Chromosomes, Artificial, Yeast; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D015723:Gene Library; D017343:Genes, Plant; D005816:Genetic Complementation Test; D005822:Genetic Vectors / Q000235:genetics*; D008969:Molecular Sequence Data; D010947:Plants, Toxic; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D014026:Nicotiana / Q000235:genetics; D014170:Transformation, Genetic
D018076:DNA, Complementary