Premium
DNA rearrangement associated with the integration of T‐DNA in tobacco: an example for multiple duplications of DNA around the integration target
Author(s) -
Ohba Toshiharu,
Yoshioka Yasushi,
Machida Chiyoko,
Machida Yasunori
Publication year - 1995
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1995.07010157.x
Subject(s) - dna , biology , inverse polymerase chain reaction , dna clamp , microbiology and biotechnology , site specific recombination , nucleic acid sequence , dna polymerase ii , genetics , dna sequencing , in vitro recombination , polymerase chain reaction , gene , molecular cloning , recombination , complementary dna , nested polymerase chain reaction , reverse transcriptase , recombinase
Transferred DNA (T‐DNA) of the tumor‐inducing (Ti) plasmid is transferred from Agrobacterium tumefaciens to plant cells and is stably integrated into the plant nuclear genome. By the inverse polymerase chain reaction DNA fragments were amplified that contained the T‐DNA/plant DNA junctions from the total DNA of a transgenic tobacco plant that had a single copy of the T‐DNA in a repetitive region of its genome. A DNA fragment containing the target site was amplified from the total DNA of non‐transformed tobacco by the polymerase chain reaction using high‐stringency conditions. Comparison of the nucleotide sequence of the target site with those of the T‐DNA/plant DNA junctions revealed that various duplications of short stretches of nucleotide sequences around the target and in the incoming T‐DNA had accompanied the integration of the T‐DNA. A deletion of 16 bp at the target site was also found and the target site was similar, in terms of nucleotide sequence, to regions around the breakpoints of the T‐DNA. This finding provides a clear example of the occurrence of complex rearrangements during the integration of T‐DNA.