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How to Isolate a Gene?

comparative Essay
870 words
870 words
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How to isolate a gene? There are various techniques present for the isolation of a gene. These techniques are used to genetically engineer the organism. First step involves the selection and then isolation of the gene of interest which needs to be genetically engineered in an organism. It is known that, most of the genes are transferred into the plant organism to provide herbicide and insects tolerance. In case of animals the genes are usually of growth hormones. One of the most common technique used till date, is by using restriction enzymes to cut the DNA into fragments and then run on gel electrophoresis for separation according to their lengths. As we know that a single strand of plant DNA or animal DNA contain tens of thousands of genes, each working for the production of a specific protein essential for the growth and survival of the organism. PCR (polymerase chain reaction) can also be used for the amplification of genes segments which can be isolated through same procedure as gel electrophoresis. Selectable markers are used for the identification and The DNA Band at the correct size should be the one containing the gene, and it can then be excised from the gel. The need of an hour is to isolate individual gene and determine its function in shorter span. By using traditional techniques to figure out merely one gene is very time consuming method. To overcome this, new procedure that enabled the scientists to identify specific genes in a matter of months, not years. This technique is known as transcript-based cloning. The bigger impact of this technique is on plants with large and complex genomes, including most crop species. The transcript based cloning technique specifically was applied on the DNA of Medicago truncatula a ... ... middle of paper ... ...ish a symbiotic relationship with nitrogen-fixing bacteria. Using microarray gene-chip technology, we monitor RNA levels produced by 10,000 genes in both normal and mutant plants. "In the mutant plants, we found one gene, called DMI3, which produced extremely low levels of RNA. The normal version of the DMI3 gene produces a protein that is remarkably similar to tobacco plant proteins that are known to modulate their behaviors in response to calcium." This finding led to conclude that the DMI3 gene may play an important role in the plant's response to calcium oscillations. It is necessary for Rhizobial and Mycorrhizal interactions and encodes a calcium and calmodulin-dependent kinase. In the process of working on nitrogen fixation, we have discovered a general method for identifying important plant genes that is fast and may be applicable to almost any plant species.

In this essay, the author

  • Explains the various techniques for the isolation of a gene, which are used to genetically engineer the organism.
  • Explains that restriction enzymes cut the dna into fragments and run on gel electrophoresis for separation according to their length. pcr (polymerase chain reaction) can also be used for amplification of genes segments.
  • Explains that the transcript-based cloning technique was applied on the dna of medicago truncatula, a member of the legume family.
  • Describes the process of destroying the maximum number of plants seed through radiation and then growing those exposed seeds in laboratory. the mutant gene is developed or raised as a result of this physical mutation.
  • Explains that if anyone is interested in finding a gene responsible for normal root growth, he would look for mutant plants with defective roots and conduct an exhaustive analysis of the plant's dna until they located the defective gene.
  • Explains that in healthy plant and animal cells, genes are responsible for the production of proteins. mutated genes carry faulty instructions that produce defective copies of rna.
  • Explains that the pnas experiment used transcript-based cloning to identify a plant gene that plays an important role in the production of usable nitrogen.
  • Explains that soil-dwelling bacteria transform atmospheric nitrogen into a compound that plants can absorb in their roots and then convert into proteins. animals get their primary source of nitrogen from plants, which make bacterial nitrogen fixation essential to all animal and human.
  • Explains that identifying genes that allow plants to establish beneficial symbiosis for nitrogen fixation is a key to sustainable agriculture.
  • Compares normal medicago plants with a mutant version raised in the lab. this mutant exhibited calcium spiking behavior but unlike their counterparts, they were unable to establish symbiotic relationships with nitrogen-fixing bacteria.
  • Explains that using microarray gene-chip technology, they monitor rna levels produced by 10,000 genes in both normal and mutant plants.
  • Concludes that the dmi3 gene may play an important role in the plant's response to calcium oscillations.
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