Mutation can be a result of damage in the genetic material through exposure to mutagens such as chemical agents, or physical agents some examples are ultraviolet rays, transposons, and ethidium bromide. (Campbell, 2005) Mutant plant cells accumulating amino acids are plant cells containing variant DNA compared to all the other normal cells.
Since the identity of the amino acids being produced is unknown, as well as the quantity of these amino acids produced by normal and mutated cells, the first step to take is the quantification and identification of amino acids in tissue culture. Immunofluorescence is an efficient method of detection of amino acids for tissue cultured cells. Through amino acid recognition segregation of functional cell types is possible, consequently, identifying the mutant cells. (Signature Immunologics, 2000) Immunofluorescence is the process by which fluorescent dyes are used for labeling antibodies to demonstrate the antibodys presence in a tissue culture.
In performing this, with Sherry Laboratory as a source, modified 30 mm polystyrene dishes are used. One shall drill out a circle on the bottom of the dish and attach a glass coverslip onto the drilled hole. A PBS formulation as vehicle shall be used for cultured cells due to its characteristic lack of embedding matrix. Liquid exchanges should not remove the primary cell specimen through the process of fixation to coverslipping. Mild fluid transfer is adequate for washes. The well created earlier should not dry out and must continuously be filled. Triton X-100 (0.1) is added to the primary antiserum diluent to improve antibody penetration. Culture dishes should be labeled with fluorophores and coverslipped in an anti-fade mountant. The dishes shall, then, be stored under optimal conditions for fluorophores (FITC 0C, TRITC 4C) (Signature Immunologics, 2000) Stored samples can, then, be viewed for observation through immunofluorescence microscopy.
After the selection of mutant plant cells accumulating amino acids, one is now able to use it for the production of secondary products in cell cultures and for crop improvement in the subsequent ways. In the production of secondary metabolites, scientists usually use plants as the major source of ingredients. These secondary metabolites are used in suspension cultures which serve as cell constituents which are not essential for specimens subjected to it. These secondary metabolites include alkaloids, glycosides, terpenoids and a variety of flavours and more. The manufacture of these products can be increased with human intervention. However, through mutations, such as those mutants accumulating amino acids levels higher, there is a probability that the production of secondary metabolites be increased. (Students Guide.In)
The inability of humans and animals to synthesize particular amino acids has been a subject of interest for a long time and researchers have focused on using crop plants to provide these essential amino acids for human and animal consumption. It was found that crop plants have naturally occurring amino acids in them. Among these amino acids are lysine, tryptophan and methionine. With mutants accumulating amino acids at hand, the production of amino acid-enriched crops will be much easier. Through genetic engineering, the isolated mutants can be transferred to crops not demonstrating high levels of amino acid production (Galili, 2008)
If the production of crops with high levels of essential amino acids is made possible, then it would greatly help mankind. With this development, people and animals will be consuming more nutritional foods. In addition, economic growth may also be achieved through this genetic breakthrough.
Answers
I. Describe a strategy for selection of mutant plant cells accumulating amino acids
- Immunofluorescence is an efficient method of detection of amino acids for tissue cultured cells. Through amino acid recognition segregation of functional cell types is possible, consequently, identifying the mutant cells.
II. Explain the potential use of such mutants
for the production of secondary products in cell cultures
- Through mutations, such as those mutants accumulating amino acids levels higher, there is a probability that the production of secondary metabolites be increased.
b. for crop improvement
- With mutants accumulating amino acids at hand, the production of amino acid-enriched crops will be much easier. Through genetic engineering, the isolated mutants can be transferred to crops not demonstrating high levels of amino acid production.
Since the identity of the amino acids being produced is unknown, as well as the quantity of these amino acids produced by normal and mutated cells, the first step to take is the quantification and identification of amino acids in tissue culture. Immunofluorescence is an efficient method of detection of amino acids for tissue cultured cells. Through amino acid recognition segregation of functional cell types is possible, consequently, identifying the mutant cells. (Signature Immunologics, 2000) Immunofluorescence is the process by which fluorescent dyes are used for labeling antibodies to demonstrate the antibodys presence in a tissue culture.
In performing this, with Sherry Laboratory as a source, modified 30 mm polystyrene dishes are used. One shall drill out a circle on the bottom of the dish and attach a glass coverslip onto the drilled hole. A PBS formulation as vehicle shall be used for cultured cells due to its characteristic lack of embedding matrix. Liquid exchanges should not remove the primary cell specimen through the process of fixation to coverslipping. Mild fluid transfer is adequate for washes. The well created earlier should not dry out and must continuously be filled. Triton X-100 (0.1) is added to the primary antiserum diluent to improve antibody penetration. Culture dishes should be labeled with fluorophores and coverslipped in an anti-fade mountant. The dishes shall, then, be stored under optimal conditions for fluorophores (FITC 0C, TRITC 4C) (Signature Immunologics, 2000) Stored samples can, then, be viewed for observation through immunofluorescence microscopy.
After the selection of mutant plant cells accumulating amino acids, one is now able to use it for the production of secondary products in cell cultures and for crop improvement in the subsequent ways. In the production of secondary metabolites, scientists usually use plants as the major source of ingredients. These secondary metabolites are used in suspension cultures which serve as cell constituents which are not essential for specimens subjected to it. These secondary metabolites include alkaloids, glycosides, terpenoids and a variety of flavours and more. The manufacture of these products can be increased with human intervention. However, through mutations, such as those mutants accumulating amino acids levels higher, there is a probability that the production of secondary metabolites be increased. (Students Guide.In)
The inability of humans and animals to synthesize particular amino acids has been a subject of interest for a long time and researchers have focused on using crop plants to provide these essential amino acids for human and animal consumption. It was found that crop plants have naturally occurring amino acids in them. Among these amino acids are lysine, tryptophan and methionine. With mutants accumulating amino acids at hand, the production of amino acid-enriched crops will be much easier. Through genetic engineering, the isolated mutants can be transferred to crops not demonstrating high levels of amino acid production (Galili, 2008)
If the production of crops with high levels of essential amino acids is made possible, then it would greatly help mankind. With this development, people and animals will be consuming more nutritional foods. In addition, economic growth may also be achieved through this genetic breakthrough.
Answers
I. Describe a strategy for selection of mutant plant cells accumulating amino acids
- Immunofluorescence is an efficient method of detection of amino acids for tissue cultured cells. Through amino acid recognition segregation of functional cell types is possible, consequently, identifying the mutant cells.
II. Explain the potential use of such mutants
for the production of secondary products in cell cultures
- Through mutations, such as those mutants accumulating amino acids levels higher, there is a probability that the production of secondary metabolites be increased.
b. for crop improvement
- With mutants accumulating amino acids at hand, the production of amino acid-enriched crops will be much easier. Through genetic engineering, the isolated mutants can be transferred to crops not demonstrating high levels of amino acid production.
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