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An enzyme is a protein-made substance that works as a catalyst in all living organisms that regulate the speed of all the chemical reactions. While accelerating the reaction process it does not get altered in the process. An enzyme is made up of chains of amino acids that are called polypeptide chains. The sequence of chains of amino acids defines the folding patterns of the structure of the protein. Enzymes regulate every aspect of metabolism in living organisms. It involves food digestion where large nutrients such as carbs, fats, and proteins get broken down into smaller pieces. An enzyme interacts with only one type of substance that is called substrate for the catalysation of a reaction.
What is genetic engineering?
Genetic engineering is a type of method where the technology of recombinant DNA (r-DNA) is used to deflect the genetic characteristics of a living organism. This genetic engineering includes the manipulation of genes that is done directly. A gene from a different species is attached to a genome of an organism to get an intended phenotype. Genetic engineering is often known as genetic modification.
Genetic engineering or genetic modification has different applications to medicines and drugs that involve gene therapy, the creation of model organisms that imitate human conditions, the manufacturing of drugs, and others. The primary use of genetic engineering was to develop human insulin inside bacteria. It is also used to develop models of animals of human diseases. The mouse is the most widely used animal for genetic engineering.
Enzymes used in genetic engineering
Different “enzymes are used in genetic engineering”. The most used enzymes are DNA polymerase, Reverse transcriptase, DNA ligase, restriction endonuclease, alkaline phosphatase. DNA polymerase functions in duplicating the content of DNA of a cell at the time of cell division. The synthesisation of DNA is the primary function of the enzyme.
Alkaline phosphatase functions to remove the single phosphate group from the 5’ ends.
The reverse transcriptase functions in generating complementary DNA from a template of RNA through the process of reverse transcription. Ligase enzymes are used in the process of joining and restriction enzymes are used to cut. Restriction enzymes can cut DNA in two various ways to yield blunt ends (cutting at the asymmetrical position and cutting at opposite sides of DNA).
A restriction endonuclease, an enzyme that is separated from bacteria that reap off molecules of DNA at certain sequences. Restriction enzymes are a type under endonuclease enzymes. These enzymes are divided into five different types (Tpe I, II, III, IV, V). Each of them is structurally different. “DNA ligase enzyme is used in genetic engineering” is broadly used for the joining of DNA strands.
The reaction of ligation or joining is regulated by different key factors such as concentration, ph, temperature, and others. “Alkaline phosphatase” is used for the modification of DNA. It removes 5′ phosphate groups from Single-stranded DNA (ss-DNA) as well as double-stranded DNA (ds-DNA). Also, it helps in preventing self-ligation.
Define primosome
Primosomes can be defined as combinations of enzymes and different proteins that are connected with DNA for the onset of replication of DNA. The main element for primosomes is the primase enzyme. This primase enzyme is accountable for the synthesis of Short RNA single strand that works as primers for the DNA polymerase. Along with the primase enzyme, it also has another enzyme that is the helicase enzyme which uncoils the DNA double strands for the replication purpose.
Components of primosome
Primosome is a complex of proteins that is accountable for the creation of RNA primers during the replication of DNA. Components of the primosome are seven different proteins such as DnaB helicase, DnaC helicase assistant, DnaT, PriA, DnaG primase, PriB, and PriC.
Primosome function
The function of the primosome is to recruit the helicase on the single-stranded DNA. It is a kind of nucleoprotein that helps in the activation of the DNA replication fork. At every fork of replication, the primosome is used. The complex is formed by the PriB, PriC, and PriA, once it binds to the DNA. After the binding, the helicase complex of DnaB-DnaC gets attached to the DnaT. This structure is called a pre-primosome. After that, DnaG gets attached to the pre-primosome complex. The primosome binds 1-10 nucleotides of RNA to the DNA single-strand and forms a DNA-RNA hybrid. This RNA sequence is utilized as a primer to start “DNA polymerase III”. The bases of RNA get replaced by DNA bases with the help of RNase H. After that DAN ligase joins both ends.
Conclusion
The “enzymes” is an important chapter in “molecular biology”. Different “enzymes are used in genetic engineering” for different purposes. A huge part of “molecular biology” involves the “enzyme” topic. For the understanding of this topic, immense study about this chapter needs to be required. In this section overview of the “enzymes” and “enzymes used in genetic engineering” is discussed in terms of their types, utilisation.
