1022-7954/05/4103- © 2005 Pleiades Publishing, Inc.
Russian Journal of Genetics, Vol. 41, No. 3, 2005, pp. 337–338. Translated from Genetika, Vol. 41, No. 3, 2005, pp. 427–429.
Original Russian Text Copyright © 2005 by Ogarkova, Tomilov, Tomilova, Tarasov.
Among current approaches to studying the structure
and function of the
which was completely sequenced in 2000, there are
such efﬁcient methods as agrobacterial transformation
making possible mass production of insertion mutants,
gene cloning, marking of T-DNA insertions, and com-
puter-assisted analysis of DNA sequences, which provides
information on the molecular structure of genes and their
protein products. In recent years, the expanded collections
mutants successfully used for studying gene
functions were obtained by the method of insertion
mutagenesis [1–5]. On the other hand, using the present-
day computer databases [http//kazusa.or.jp/, http//
http//www.ncbi.nlm.nih.gov/] a gene can be identiﬁed
from a short known DNA sequence near insertion. This
approach was used in the present study to identify the
gene with insertion leading to changes in morphology
We used the collection of inser-
tion morphological mutants of
Köln, obtained by agrobacterial transformation of ger-
minating seeds and using the binary vector system
pLD3 . Earlier, we described in detail the media, the
cultivation conditions, selection of transformed plants
and testing for their transgenic nature, histochemical
analysis of the reporter gene of
isolation and analysis of DNA from plant tissues, con-
ditions and primers for PCR used to detect the trans-
formed plant T-DNA, and determining the number of T-
DNA insertions .
Isolation and characterization of the mutant.
recessive and lethal mutant of
represented by line 281 in the collection.
After cocultivation with agrobacteria, the ﬁrst gen-
eration (T1) seeds were germinated aseptically in soil.
The transgenic plant selection was brought about in the
following generation (T2) . As a result, a kanamy-
cin-resistant plant with normal morphology was
obtained, the cells of which exhibited GUS activity,
whereas PCR analysis conﬁrmed the presence of T-
DNA in preparations of genomic DNA of the plant.
Southern blot hybridization showed that the mutant
genome of line 281 carried a single insertion (data not
Identification of the Gene Whose Mutation
Leads to Hypocoptyl Tropism in
O. A. Ogarkova, A. A. Tomilov, N. B. Tomilova, and V. A. Tarasov
Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, 119991 Russia;
fax: (095)135-43-04; e-mail: firstname.lastname@example.org
Received July 13, 2004
—Genetic and molecular analysis of a mutant of
with bended hypocoptyl from
a previously obtained collection of insertion mutants is presented. The examined mutation was shown to be
recessive and based on a single insertion of pLD3 vector T-region into the
TAIL-PCR analysis of a DNA region adjacent to the left border of the insertion revealed a putative site of T-
DNA insertion, the 609-bp
gene from chromosome 1 represented by a single exon.
40 41 46 50 51 6021 29 30 31
The cloned nucleotide sequence of the genomic DNA from the mutant line 281 (bold and italicized), which is adjacent to the left
border of T-DNA insertion.