Gene cloning is the process by which molecular biology tools are used in the development of identical copies of a specific gene. The process involves the seclusion of DNA sequence and introducing it to a vector for proliferation without changing the native DNA sequence. The method of secluding the DNA is referred to as molecular cloning (Ashwani, Murugan, Balamurugan, & Sathiskumar, 2016). The clones obtained after seclusion can be used to generate many copies of DNA for analyzing the gene sequence of a protein and also modifying it in vitro to transform the function of the protein (Ashwari et al., 2016).
Scientists have developed various techniques for molecular cloning. One of the traditional methods developed in the 20th century was the restriction enzyme cutting and litigation of the recombinant DNA assembly (Ashwani et al., 2016). Other contemporary techniques have been developed such as the conventional cloning and gateway cloning. Traditional cloning involves the isolation of the target DNA fragment, ligation of inserts into the selected vector, modification of recombinant plasmids for propagation, and screening of the hosts comprising the desired plasmid (Ashwani et al., 2016). Gateway cloning is widely accepted due to its ability to arrange DNA fragments in an advanced order, orientation, and reading frame notwithstanding the sequence of the DNA.
Molecular cloning has applications in plant and clinical microbiology. The simplicity, cost-effectiveness, reliability, and rapidity of molecular cloning have become a crucial tool in clinical microbiology. Clinical microbiology involved the isolation of pathogens and categorizing of the isolates according to their respective human infectious disease (Sharma, Mishra, Duraisamy, & Mehraj, 2014). The technique was limited and practiced in a few places in the world. However, the development of microbiology has brought about clarity and speed in microbiological diagnosis. In clinical microbiology, scientists are utilizing molecular cloning to identify polymicrobial diseases (Sharma et al., 2014). Molecular cloning has also helped in the creation of recombinant antigens that are now used in the development of serological screening assays for the detection, quantification, and characterization of hepatitis C virus (HCV). Besides, molecular cloning is also utilized in the creation of recombinant vaccines. These vaccines are cost-effective and are abundantly available. The developed vaccines can be used for various infectious diseases such as cholera, hepatitis B, HIV, and malaria (Sharma et al., 2014). Molecular cloning in clinical microbiology has other applications such as the development of diagnostic probes, antimicrobial peptides, and recombinant cytokines.
In plant microbiology, molecular cloning is used in the isolation, characterization, and expression analysis of flavonoids, which are the key enzymes in muscadine grape development (Hall, Ananga, Georgiev, Ochieng, Cebert, & Tsolova, 2015). The study by Hall et al. (2015) managed to elaborate and display the isolation, cloning, and characterization of VrF3H gene from muscadine grapes. Molecular cloning helped in the understanding of the flavonoid pathway in muscadine grapes. Equally important, molecular cloning has also been used to explain the evolution of plant nonsymbiotic hemoglobins (nsHbs). A study by Garrocho-Villegas and Areedondo-Peter (2008) showed the molecular cloning and characterization of a nonsymbiotic gene from a moss plant. The cloning process helped the researchers to determine that moss nsHbs have been known for a long time and the size of the nsHbs have been decreasing over time due to the lack of amino acids (Garrocho-Villegas, 2008).
In conclusion, molecular cloning, which is the process of isolating DNA sequence proves to be a reliable method in both plant and clinical microbiology. The technique has helped in the analysis of plant evolution and the extraction process of muscadine grapes. In clinical microbiology, molecular cloning has played a significant role in the development of vaccines, identification of various diseases, and the development of different diagnostic probes.
Ashwini, M., Murugan, S., Balamurugan, S., & Sathishkumar, R. (2016). Advances in molecular cloning. Molecular Biology, 50 (1), 1-6.
Garrocho-Villegas, V., & Arredondo-Peter, R. (2008). Molecular cloning and characterization of a moss (ceratodon purpureus) nonsymbiotic hemoglobin provides insight into the early evolution of plant nonsymbiotic hemoglobins. Mol. Biol. Evol, 25(7), 14821487. doi:10.1093/molbev/msn096
Hall, J., Ananga, A., Georgiv, V., Ochieng, J., Cebert, E., & Tsolova, V. (2015). Molecular cloning, characterization, and expression analysis of flavanone 3-hydroxylase (f3h) gene during muscadine grape berry development. J Biotechnol Biomater 5(2), 1-7. http://dx.doi.org/10.4172/2155-952X.1000180
Sharma, K., Mishra, A., Duarisamy, G., & Mehraj, V. (2014). Advances and applications of molecular cloning in clinical microbiology. Biotechnology and Genetic Engineering Reviews, 30(1), 65-78. DOI: 10.1080/02648725.2014.921501
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