Characterization of “Landmark Protein” from Rhizobium isolates of pigeon pea (Cajanus cajan (L.) Millsp.) collected from acidicsoils of the State Of Jharkhand by utilizing immunological approach
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Date
2020
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Birsa Agricultural University, Ranchi
Abstract
Soil acidity occurs when there is a buildup of acid in soil.Acidic soil in Jharkhand occupies
approximately 90 percent area of the land. Most leguminous plant requires neutral or slightly
acidic soil for growth, especially when depending on symbiotic nitrogen fixation.
Pigeon pea is legume, soil fertility through biological nitrogen fixation.It is reported to
contribute approximately 40 kg N ha-1 soil acidity is one of the serious problem which affect
growth of Rhizobia in the soil of Jharkhand.Rhizobium is a Gram negative, soil living bacteria.
Rhizobium provides organic nitrogenous compound to the plant and plant provide ompound
made by photosynthesis, this relation is known as symbiosis
Two-dimensional electrophoresis (2-D electrophoresis) is a powerful and widely
usedmethod for the analysis of complex protein mixtures extracted from cells, tissues, or
otherbiological samples. This technique sort’s protein according to two independent properties
intwo discrete steps: the first-dimension step, isoelectric focusing (IEF), separates
proteinsaccording to their isoelectric points (pI); the second-dimension step, SDSpolyacrylamide
gelelectrophoresis (SDS-PAGE), separates proteins according to their molecular
weights. Eachspot on the resulting two-dimensional array corresponds to a single protein species
in thesample. MALDI measures the mass of peptides derived from a trypsinized parent protein
and generates a list of experimental peptide masses, often referred to as „mass fingerprints‟
During MALDI-TOF analysis, the m/z ratio of an ion is measured by determining the time
required for it to travel the length of the flight tube
A method for the specific histochemical demonstration of antibody in cells and parts of
cells is described. It consists of carrying out a two stage immunological reaction on frozen
sections of tissues: (a) allowing reaction between antibody in the tissue and dilute antigen
applied in vitro, and (b) the detection of those areas where this antigen has been specifically
absorbed by means of a precipitin reaction carried out with fluorescein-labelled antibody.
Western blotting (or immunoblotting) is a widely used method to detect proteins as well as post
translational modification on proteins, using antibody based on probes to obtain specific
information about target proteins from complex samples. It can provide semi-quantitative or
quantitative data about the target protein in a simple or complex biological sample.
The aim of my work is to“Characterization of “Landmark Protein” from Rhizobium
isolates of pigeon pea (Cajanus cajan (L.) Millsp.) collected from acidic-soils of the State Of
Jharkhand by utilizing immunological approach”
Keeping in mind the above background, the research work was carried out with the following
broad objectives:
Protein samples processing for SDS-PAGE, for Rhizobium isolates.
SDS-PAGE profiling of protein.
Generation of antibody profiles by ELISA and Western Blot analysis of proteins
Description
Characterization of “Landmark Protein” from Rhizobium isolates
of pigeon pea (Cajanus cajan (L.) Millsp.) collected from acidicsoils
of the State Of Jharkhand by utilizing immunological
approach