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What is Genetic engineering ? How is a beneficial gene amplified through PCR technique ? Explain.
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Genetic engineering technique is used to modify the genetic material of an organism. The organism is called as genetically modified organism. GMO stands for genetically modified organisms. Plants, animals or any other microorganisms which have a new type of genetic material which have been modified through genetic engineering are termed genetically modified organisms.The gene may be inserted into the host genome. The gene is first isolated from the cell. The desired gene is restricted with the help of specific restriction enzymes. These enzymes are also used to cleave the plasmid vector. The desired gene is inserted in the plasmid vector with the help of enzyme ligase. This results in the formation of recombinant DNA. Many different segments of DNA can be attached in the vector. The plasmids are cloned in the bacterial cells by using molecular cloning methods.
The polymerase chain reaction (PCR) is a technology in molecular biology used to amplify a single copy of DNA to millions of copies of a particular DNA sequence. It consists of a series of 20-40 repeated temperature changes, called cycles. Denaturation step: The reaction is heated to 94-98o C for 20-30 seconds. It causes disrupting the hydrogen bonds between complementary bases, yielding single-stranded DNA molecules.Annealing step: The reaction temperature is lowered to 50-65o C for 20-40 seconds allowing annealing of the primers to the single-stranded DNA template. Extension/elongation step: Taq polymerase is the enzyme used and the reaction is carried out at 72o C. Taq DNA polymerase synthesizes a new DNA strand complementary to the DNA template strand by adding dNTPs. This results in multiple copies of the desired gene which can be used for genetic engineering. There are many applications of PCR:- This technique is commonly used in DNA finger printing method for the amplification of the DNA.
- Helps in the diagnosis of mutation and genetic disorders.
- Also used in the diagnosis of infectious agents.
The gene may be inserted into the host genome. The gene is first isolated from the cell. The desired gene is restricted with the help of specific restriction enzymes. These enzymes are also used to cleave the plasmid vector. The desired gene is inserted in the plasmid vector with the help of enzyme ligase. This results in the formation of recombinant DNA. Many different segments of DNA can be attached in the vector. The plasmids are cloned in the bacterial cells by using molecular cloning methods.
The polymerase chain reaction (PCR) is a technology in molecular biology used to amplify a single copy of DNA to millions of copies of a particular DNA sequence. It consists of a series of 20-40 repeated temperature changes, called cycles.
Denaturation step: The reaction is heated to 94-98o C for 20-30 seconds. It causes disrupting the hydrogen bonds between complementary bases, yielding single-stranded DNA molecules.
Annealing step: The reaction temperature is lowered to 50-65o C for 20-40 seconds allowing annealing of the primers to the single-stranded DNA template.
Extension/elongation step: Taq polymerase is the enzyme used and the reaction is carried out at 72o C. Taq DNA polymerase synthesizes a new DNA strand complementary to the DNA template strand by adding dNTPs.
This results in multiple copies of the desired gene which can be used for genetic engineering.
There are many applications of PCR:
- This technique is commonly used in DNA finger printing method for the amplification of the DNA.
- Helps in the diagnosis of mutation and genetic disorders.
- Also used in the diagnosis of infectious agents.
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