Agriculture Reference
In-Depth Information
Other methods are also available to monitor gene expression and distribution of
proteins within cells. They made use of coding sequences for beta-galactosidase,
luciferase, or bacterial luciferase for the formation of fused proteins (Chalfie
et al.
1994
). The requirement of exogenously adding of substrate or cofactors limits
their application in living tissues.
Intracellular detection of GFP requires only irradiation by UV providing an easy
means for the detection of gene expression and protein localization in living cells
(eukaryotic and prokaryotic) (Chalfie et al.
1994
; Dhandayuthapani et al.
1995
;Bo
Andersen et al.
1998
). Another advantage of GFP is related to the fact that this does
not interfere with cell growth and cell physiology. For the use of some proteins as a
marker for gene transfer, the characteristics of the cell being studied and factors
such as the utilization of fixed compounds should be taken into account. The use of
fixed favor maintenance of the characteristics of cells for the study, however, can
directly affect the fluorescence of the protein (Chalfie and Kain
1998
). The choice
of methods for fluorescence quantification is primordial to use GFP as a marker
protein. Niedenthal et al. (
1996
) have reported the most common and fastest
methods to determine GFP behavior in cells, which include methods such as
epifluorescence microscopy, fluorometry, and flow cytometry.
Crystal structures of high-resolution GFP offer unprecedented opportunities for
understanding and manipulating the relationship between the structure and function
of protein spectroscopy. GFP has become a well-known marker system for gene
expression and protein detection in intact target cells and organisms, opening new
perspectives in physiological indicators, biosensors, and gene markers (Tsien
1998
).
New method of gene transformation in plants with nanoparticles is the new area in
plant biotechnology (Fig.
10.1
).
Plant tissue culture technique is an essential tool in agriculture and medicinal
plant research. It depends on maintaining plant cells in controlled aseptic conditions
on basal nutrient medium with appropriate hormonal concentration. The plant cell
culture can be sustained as a callus (mass of undifferentiated cells) for an extended
period of time or can be regenerated into whole plant. For the production of
secondary plant metabolites and regeneration of plant with improved nutritional
quality, higher yields, and tolerance against biotic and abiotic stresses, newer genes
need to be introduced in the preexisting plants with normal physico- and phyto-
chemical properties (Dodds and Roberts
1990
; Bhojwani and Razdan
1996
; Ohadi
Fig. 10.1 The new method of transformation of plants with transgenes encapsulated in
nanocarriers is relatively the new area in plant biotechnology
