Agriculture Reference
In-Depth Information
10.3.5 Temperature-Mediated Gene Transfer
Raising the temperature enhances gene transfer that is successfully adopted and
confirmed to animal cell transfection process (Kenel et al.
2010
). It seems from the
literature that at a temperature above 37
C, the gene transfer tendency increases
and further increase up to 43
C with a certain period of time provides greater
transient transfection. This was confirmed by the work on interleukin-2 and swine
growth hormone expressions using indirect ELISA (Dillen et al.
1997
; Baron
et al.
2001
). Dillen et al. (
1997
) demonstrated the effect of temperature (15-
29
C) on
Agrobacterium
-mediated gene transfer in plants. Baron et al. (
2001
)
reported the effect of temperature in biotransformation process with
A. tumefaciens
involved in cocultivation of
Phaseolus acutifolius
and further in
Nicotiana tabacum
biotransformation. In both the situations, the level of transient
uidA
expression
decreased notably when the temperature was raised above 22
C, further lowered
down in temperature at 27
C, and was undetectable at 29
C (Baron et al.
2001
; Iba
2002
; Rakoczy-trojanowska
2002
). Commonly used conventional methods of bio-
transformation discussed here are summarized in Table 10.
1
along with their
advantages, mechanisms of gene transfer, and limitations.
10.3.6 Microinjection
In case of microinjection, DNA is directly injected into plant nucleus or cytoplasm,
using a fine tip of 0.5-1.0
ΚΌ
m in diameter made up of glass needle or micropipette.
Table 10.1 Commonly used conventional methods of bio-transformation
Method
Advantages
Disadvantages
Viral
High efficient
Immunogenicity, carcinogenicity,
a risk of recombination in case
animal cells
Electroporation
Safe, easy, rather efficient
Need large amount DNA and has
to be optimized for every cell type
Microinjection
Easy and highly efficient
Only one cell at a time can be
transfected, no suitable for whole
organism
Gene gun
Can delivered thousand of DNA copies,
high transgen expression
Shallow penetration of DNA in
tissue
Chemical
method
Efficient
High toxicity
Nanoparticles
Highly efficient, size tuneable from 1 to
100 nm, possible to incorporate different
functional groups on single particles,
immunogenicity can be controlled
