Advertisement
4.1552.
PCR, Southern and Western blot analyses of Chi transformants
When the genomic
DNA
of transformants was used
:is
template, gene-s(wcilic
primers amplified
1.1
kb of chitinase gene in the
5
transfornlants and positi\rc control
(pCAMBIA-cah-Bor2chi).
Conversely, no such h:ind
was observed
in
the
untransformed control under identical conditions (Fig 32
A).
Southern blot analysis was carried out on all the transformants using Chi
coding sequences as probes. Three transform~nts (plant
1.
3 and 5 ) displayed one
hybridizable band and
2
transformants (plant 2 and 4) exhibited two hyhridiz;~hle
hands for Chi probe from the total DNA of transformants indicating the stable
integration of marker gene Chi in transgenic tobacco plants (Fig. 328).
Transgenic tobacco plants were studied for the expression of chitinace. Barley
chitinase antiserum detected expression of
35
kDa polypeptide in all the
5
transformants, whereas such expression was absent in the untransfonned control plant
(Fig. 32C).
Advertisement
