Recombinant DNA Technology and Molecular Basis of Cancer

Recombinant DNA Technology and Molecular Basis of Cancer

Recombinant DNA Technology

Recombinant DNA molecules are DNA sequences that result from molecular replications hence combining genetic materials found in biological organisms together. The process of combining DNA molecules is possible due to the fact that all organisms share the same chemical structure, which only differs in nucleotides. Researchers are able to identify the causative aspects of cancer in molecular structuring. Cancer is a genetic disease that becomes aggressive overtime and disrupts the body tissues and organs needed for organism survival. Recombinant DNA is applied widely in biotechnology and basic research in identifying and mapping affected genes and even determines the function of each.

Understanding Techniques of DNA Digestion

  • Restriction Enzymes

Restriction enzymes slash DNA into nucleotide sequences identified as restriction sites. Such types of enzymes are found in archaea and bacteria, which are known to release a certain defense mechanism for attacking viruses. The selective enzyme in the restriction sites combats foreign DNA through the restriction process. Through a modification enzyme known as methylase, the host DNA is methylated in order to protect the activities of restriction enzymes (Glick, & Pasternak, 2003). Two incisions are made in the process of restricting the DNA. Restriction enzymes play vital roles in recombinant DNA molecules constructions thus restriction enzymes can be used for DNA modification and manipulation processes.

  • The Ligase Reaction

The ligase enzyme catalyzes hydrolysis and lipid formation. Ligase repairs DNA molecules from a single-stranded nature into double strands in the replication process. DNA ligase purifies gene replication in order to enable the DNA molecules to join. The DNA ligase requires a phosphodiester bond (Brown, 2006) in order to enable it repair DNA fully. DNA ligase mechanism forms two covalent phosphodiester bonds between the contributor and the receiving molecules. DNA ligases are widely used in contemporary molecular researches in order to produce recombinant DNA.

  • Electrophoresis Techniques for DNA Analysis

Electrophoresis is a DNA technique used in separating proteins in accordance to size and charge. The technique is used in molecular biology whereby DNA and RNA portions are estimated in size and separated from proteins by charge. This is achieved through separating nucleic acid molecules through the application of electric field that move the negatively charged ions into an agarose matrix. The longer molecules move slowly but the shorter move faster through the gel pores in a process termed as sieving. The charge separates proteins since the gel pores are larger and therefore unable to sieve proteins.

  • Nucleic Acid Hybridization Methods

DNA hybridization technique measures the extent of genetic connections between DNA pool sequences. It determines the distance of two species of genetic strands in nucleic acids. The connection between the two allows diverse species to be positioned in a phylogenetic tree thus making it possible in carrying out molecular processes systematically. Under normal conditions, the DNA binds two strands together. The annealing technique is used for diversity reduction thus obtaining hybrids, which are most preferred in terms of energy aspects (Klug, & Cummings, 2003).

  • Nucleotide Sequencing Techniques

Nucleotides are molecules that when combined form DNA and RNA structures. They play vital roles in metabolism and serve as chemical energy sources known as adenosine and guanosine triphosphates. They are incorporated in vital enzymatic reaction cofactors. The enzymatic techniques of DNA sequencing are based on partial digestion of duplex DNA. After a systematic DNA cleavage with enzyme restrictions, gel electrophoresis separates the labeled products and the entire process begins with unrestricted DNA followed by gel electrophoresis.

Approaches used for DNA Amplification, Particularly in the Polymerase Chain Reaction

Various approaches are employed in DNA amplification with the Polymerase Chain Reaction (PCR) being the principle one. The PCR amplifies one or many DNA copies across diverse magnitudes thus generating colossal copies of a particular DNA sequence. The technique relies on thermal cycling whereby DNA enzyme replication and DNA melting are heated and cooled down. Many PCR applications employ the thermal cycling technique for synthesizing DNA polymerase of the targeted DNA. The amplification specificity depends on the degree in which primers recognize and bind sequences other than the intended DNA sequences. PCR thermal cycling separates two DNA strands at high temperatures through DNA melting processes but at lower temperatures, the DNA polymerase amplifies the target DNA.

Ways that Recombinant DNA Techniques are Applied in Molecular Analysis and Diagnosis of Genetic Defects

Recombinant DNA techniques may be applied in molecular analysis through use of two molecular approaches; direct and indirect study detection by linkage analysis. The indirect analysis diagnoses genetic defects whereby the affected structural gene is unknown. The analysis is done through the polymorphisms analysis technique whereby the diagnostic technique restricts fragment length polymorphisms. The probability of recombination events depends on their distance (Glick, & Pasternak, 2003). Another way that may be applied is the use of direct detection of a mutated gene but this is employed in case the gene is known and the structure well defined. The approach is sensitive for diseases whereby molecular defects indicate slight inconsistencies. It is used in combination with restrictions fragment length polymorphisms analysis.

The Clinical Application of PCR Technology in the Diagnostic Laboratory

PCR is one of the vitro techniques that have been used for many years in the laboratory diagnosis of DNA amplification. It is an essential technique used in clinical practices for simplifying the DNA replication process, which occurs during cell distribution. The PCR consists of thermal cycling for target DNA, primer annealing and annealed premier extension of DNA polymerase (Sandhu, 2010). PRC applications include virology regulation whereby it has been practical in resisting testing genotyping as well as viral load modification. More rapid diagnosis has been advanced for fungal infections through the antibiotic application. Other application areas include epidemiology and control of infection achieved with use of molecular methods.

Molecular Basis of Cancer

The Difference between a Proto-Oncogene and an Oncogene

The difference between the two is traced to their body functions. A proto-oncogene is a normal gene that has diverse functions in replenishing body cells whereas an oncogene is a gene that causes cancer and can actively promote tumor growth. A proto-oncogene can develop into an oncogene in case the environmental conditions for its modification arise. A proto-oncogene becomes an oncogene when it undergoes the mutilation process. Another difference is that the proto-oncogene gene facilitates growth of development cells for in human beings whereas the oncogene promotes the development of abnormal genes that lead to cancer. Additionally, the proto-oncogene genes are responsible in the synthesis of proteins, which are vital in stimulating distribution of body cells but oncogene genes do not thus proto-oncogenes fight to control oncogenes from affecting body tissues.

Types of Protein Coding Genes That Are Usually Oncogenic

Various types of protein coding genes are usually oncogenic. First, receptor tyrosine kinases comprising of protein bases are quite oncogenic. These include epidermal growth factor receptor and vascular endothelial growth factor receptor proteins. Tyrosine kinases target proteins and cause cancer through affecting the receptor permanently. Secondly, regulatory GTPases is another type of protein coding gene that is highly oncogenic. For example the Ras protein which is a small GTPase, which hydrolyses GTP into GDP and phosphate. Lastly, the transcription factor is another protein-coding gene capable of becoming oncogenic. An example is the myc gene, which regulates gene transcription that induces cell propagations (Tortora, Funke, & Case, 2005).

Involvements of DNA Mismatch Repair Genes in Colon Cancer

DNA Mismatch Repair (MMR) genes are vital in maintaining genomic stability. The MMR primary function is to eliminate the single-base mismatch that occurs during DNA replication. Another function is to maintain DNA reliability during the duplication process. The involvement of DNA MMR in colon cancer is centered on its ability to maintain genomic stability. MMR increases DNA replication through identifying and exercising a single-base mismatch that arises during replication. Therefore, the MMR system damages DNA surveillance functions thus preventing incorrect single-base bearing of DNA polymerase. This may lead to the accumulation of mutations once cells conjoin with DNA polymerase contributing to cancer initiation. This requires different types of proteins to aid in the completion of MMR system. In colon cancer, hypermethylation is one type of protein gene that results in a transcriptional format that has been observed in diverse mutations. However, the stage in which this mutation occurs involves a wild-type MMR gene that has not been clearly defined. Few genes target MMR and these largely contribute to gastrointestinal tumors (Mendelsohn, 2001). Note that, the genes target the transformation of growth factors known as receptor II, phosphates, growth factor II receptor and cell cycle regulator for atypical growths that lead to cancer.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

References

Brown, T. A. (2006). Gene cloning and DNA analysis: An introduction. Oxford, UK: Blackwell Pub.

Glick, B. R., & Pasternak, J. J. (2003). Molecular biotechnology: Principles and applications of

 recombinant DNA. Washington, DC: ASM Press.

Klug, W., & Cummings, M. (2003). Concepts of genetics. Upper Saddle River, NJ: Prentice

Hall.

Mendelsohn, J. (2001). The molecular basis of cancer. Philadelphia, PA: Saunders.

Sandhu, S. S. (2010). Recombinant DNA technology. New Delhi, India: I.K. International Pub. House.

Tortora, G. J., Funke, B. R., & Case, C. L. (2005). Microbiology: An introduction. San

Francisco, CA: Benjamins Cummings.

 

Recombinant DNA Technology

Recombinant DNA molecules are DNA sequences that result from molecular replications hence combining genetic materials found in biological organisms together. The process of combining DNA molecules is possible due to the fact that all organisms share the same chemical structure, which only differs in nucleotides. Researchers are able to identify the causative aspects of cancer in molecular structuring. Cancer is a genetic disease that becomes aggressive overtime and disrupts the body tissues and organs needed for organism survival. Recombinant DNA is applied widely in biotechnology and basic research in identifying and mapping affected genes and even determines the function of each.

Understanding Techniques of DNA Digestion

  • Restriction Enzymes

Restriction enzymes slash DNA into nucleotide sequences identified as restriction sites. Such types of enzymes are found in archaea and bacteria, which are known to release a certain defense mechanism for attacking viruses. The selective enzyme in the restriction sites combats foreign DNA through the restriction process. Through a modification enzyme known as methylase, the host DNA is methylated in order to protect the activities of restriction enzymes (Glick, & Pasternak, 2003). Two incisions are made in the process of restricting the DNA. Restriction enzymes play vital roles in recombinant DNA molecules constructions thus restriction enzymes can be used for DNA modification and manipulation processes.

  • The Ligase Reaction

The ligase enzyme catalyzes hydrolysis and lipid formation. Ligase repairs DNA molecules from a single-stranded nature into double strands in the replication process. DNA ligase purifies gene replication in order to enable the DNA molecules to join. The DNA ligase requires a phosphodiester bond (Brown, 2006) in order to enable it repair DNA fully. DNA ligase mechanism forms two covalent phosphodiester bonds between the contributor and the receiving molecules. DNA ligases are widely used in contemporary molecular researches in order to produce recombinant DNA.

  • Electrophoresis Techniques for DNA Analysis

Electrophoresis is a DNA technique used in separating proteins in accordance to size and charge. The technique is used in molecular biology whereby DNA and RNA portions are estimated in size and separated from proteins by charge. This is achieved through separating nucleic acid molecules through the application of electric field that move the negatively charged ions into an agarose matrix. The longer molecules move slowly but the shorter move faster through the gel pores in a process termed as sieving. The charge separates proteins since the gel pores are larger and therefore unable to sieve proteins.

  • Nucleic Acid Hybridization Methods

DNA hybridization technique measures the extent of genetic connections between DNA pool sequences. It determines the distance of two species of genetic strands in nucleic acids. The connection between the two allows diverse species to be positioned in a phylogenetic tree thus making it possible in carrying out molecular processes systematically. Under normal conditions, the DNA binds two strands together. The annealing technique is used for diversity reduction thus obtaining hybrids, which are most preferred in terms of energy aspects (Klug, & Cummings, 2003).

  • Nucleotide Sequencing Techniques

Nucleotides are molecules that when combined form DNA and RNA structures. They play vital roles in metabolism and serve as chemical energy sources known as adenosine and guanosine triphosphates. They are incorporated in vital enzymatic reaction cofactors. The enzymatic techniques of DNA sequencing are based on partial digestion of duplex DNA. After a systematic DNA cleavage with enzyme restrictions, gel electrophoresis separates the labeled products and the entire process begins with unrestricted DNA followed by gel electrophoresis.

Approaches used for DNA Amplification, Particularly in the Polymerase Chain Reaction

Various approaches are employed in DNA amplification with the Polymerase Chain Reaction (PCR) being the principle one. The PCR amplifies one or many DNA copies across diverse magnitudes thus generating colossal copies of a particular DNA sequence. The technique relies on thermal cycling whereby DNA enzyme replication and DNA melting are heated and cooled down. Many PCR applications employ the thermal cycling technique for synthesizing DNA polymerase of the targeted DNA. The amplification specificity depends on the degree in which primers recognize and bind sequences other than the intended DNA sequences. PCR thermal cycling separates two DNA strands at high temperatures through DNA melting processes but at lower temperatures, the DNA polymerase amplifies the target DNA.

Ways that Recombinant DNA Techniques are Applied in Molecular Analysis and Diagnosis of Genetic Defects

Recombinant DNA techniques may be applied in molecular analysis through use of two molecular approaches; direct and indirect study detection by linkage analysis. The indirect analysis diagnoses genetic defects whereby the affected structural gene is unknown. The analysis is done through the polymorphisms analysis technique whereby the diagnostic technique restricts fragment length polymorphisms. The probability of recombination events depends on their distance (Glick, & Pasternak, 2003). Another way that may be applied is the use of direct detection of a mutated gene but this is employed in case the gene is known and the structure well defined. The approach is sensitive for diseases whereby molecular defects indicate slight inconsistencies. It is used in combination with restrictions fragment length polymorphisms analysis.

The Clinical Application of PCR Technology in the Diagnostic Laboratory

PCR is one of the vitro techniques that have been used for many years in the laboratory diagnosis of DNA amplification. It is an essential technique used in clinical practices for simplifying the DNA replication process, which occurs during cell distribution. The PCR consists of thermal cycling for target DNA, primer annealing and annealed premier extension of DNA polymerase (Sandhu, 2010). PRC applications include virology regulation whereby it has been practical in resisting testing genotyping as well as viral load modification. More rapid diagnosis has been advanced for fungal infections through the antibiotic application. Other application areas include epidemiology and control of infection achieved with use of molecular methods.

Molecular Basis of Cancer

The Difference between a Proto-Oncogene and an Oncogene

The difference between the two is traced to their body functions. A proto-oncogene is a normal gene that has diverse functions in replenishing body cells whereas an oncogene is a gene that causes cancer and can actively promote tumor growth. A proto-oncogene can develop into an oncogene in case the environmental conditions for its modification arise. A proto-oncogene becomes an oncogene when it undergoes the mutilation process. Another difference is that the proto-oncogene gene facilitates growth of development cells for in human beings whereas the oncogene promotes the development of abnormal genes that lead to cancer. Additionally, the proto-oncogene genes are responsible in the synthesis of proteins, which are vital in stimulating distribution of body cells but oncogene genes do not thus proto-oncogenes fight to control oncogenes from affecting body tissues.

Types of Protein Coding Genes That Are Usually Oncogenic

Various types of protein coding genes are usually oncogenic. First, receptor tyrosine kinases comprising of protein bases are quite oncogenic. These include epidermal growth factor receptor and vascular endothelial growth factor receptor proteins. Tyrosine kinases target proteins and cause cancer through affecting the receptor permanently. Secondly, regulatory GTPases is another type of protein coding gene that is highly oncogenic. For example the Ras protein which is a small GTPase, which hydrolyses GTP into GDP and phosphate. Lastly, the transcription factor is another protein-coding gene capable of becoming oncogenic. An example is the myc gene, which regulates gene transcription that induces cell propagations (Tortora, Funke, & Case, 2005).

Involvements of DNA Mismatch Repair Genes in Colon Cancer

DNA Mismatch Repair (MMR) genes are vital in maintaining genomic stability. The MMR primary function is to eliminate the single-base mismatch that occurs during DNA replication. Another function is to maintain DNA reliability during the duplication process. The involvement of DNA MMR in colon cancer is centered on its ability to maintain genomic stability. MMR increases DNA replication through identifying and exercising a single-base mismatch that arises during replication. Therefore, the MMR system damages DNA surveillance functions thus preventing incorrect single-base bearing of DNA polymerase. This may lead to the accumulation of mutations once cells conjoin with DNA polymerase contributing to cancer initiation. This requires different types of proteins to aid in the completion of MMR system. In colon cancer, hypermethylation is one type of protein gene that results in a transcriptional format that has been observed in diverse mutations. However, the stage in which this mutation occurs involves a wild-type MMR gene that has not been clearly defined. Few genes target MMR and these largely contribute to gastrointestinal tumors (Mendelsohn, 2001). Note that, the genes target the transformation of growth factors known as receptor II, phosphates, growth factor II receptor and cell cycle regulator for atypical growths that lead to cancer.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

References

Brown, T. A. (2006). Gene cloning and DNA analysis: An introduction. Oxford, UK: Blackwell Pub.

Glick, B. R., & Pasternak, J. J. (2003). Molecular biotechnology: Principles and applications of

 recombinant DNA. Washington, DC: ASM Press.

Klug, W., & Cummings, M. (2003). Concepts of genetics. Upper Saddle River, NJ: Prentice

Hall.

Mendelsohn, J. (2001). The molecular basis of cancer. Philadelphia, PA: Saunders.

Sandhu, S. S. (2010). Recombinant DNA technology. New Delhi, India: I.K. International Pub. House.

Tortora, G. J., Funke, B. R., & Case, C. L. (2005). Microbiology: An introduction. San

Francisco, CA: Benjamins Cummings.

 

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