Rekombinant Dna Teknolojisi ve Kullanılan Mikroorganizmalar
Özet
Bu kitap bölümü, rekombinant DNA teknolojisinin temel prensiplerini, kullanılan başlıca mikroorganizmaların avantajları ve dezavantajları, bu teknolojinin çeşitli sektörlerdeki güncel uygulamalarını kapsamlı bir şekilde ele almayı amaçlamaktadır. Ayrıca, teknolojideki son gelişmeler, özellikle CRISPR-Cas9 gibi gen düzenleme araçları ve sentetik biyolojinin rolü üzerinde durulacak, etik ve güvenlik konularına değinilerek gelecekteki potansiyel uygulamalar ve zorluklar değerlendirilecektir. Mikroorganizmaların rekombinant DNA teknolojisindeki merkezi rolü, bu bölümün ana odak noktalarından biri olacaktır, zira bakteriler ve mayalar gibi mikroorganizmalar, genetik mühendisliği uygulamaları için vazgeçilmez konak sistemleri sunmaktadır. Bu bölüm, okuyuculara rekombinant DNA teknolojisinin karmaşık dünyasına dair derinlemesine bir bakış sunarak, bu alandaki güncel bilgi birikimini ve gelecekteki yönelimleri anlamalarına yardımcı olmayı hedeflemektedir. Ayrıca yeteri miktarda üretilemeyen bileşiklerin yüksek miktarda üretimine yönelik alternatif çalışma modelleri hakkında literatürde yapılan bazı çalışmaları (Albümin ve İnsülin gibi hayati öneme sahip proteinler ve günlük hayatımızda’da kullanabileceğimiz hidrofobin proteinler) değerlendireceğiz.
Referanslar
Nanoteknoloji. Biyoteknoloji nedir? (27.08.2025 tarihinde https://nanoteknoloji.org/biyoteknoloji-nedir. adresinden ulaşılmıştır.
Timmis K, de Vos WM, Ramos J L, et al. The contribution of microbial biotechnology to sustainable development goals. Microb Biotechnol. 2017; 10(5):984-987. doi: 10.1111/1751-7915.12818.
Hasija Y. Chapter 5. All About Bioinformatics. 2023, Pages 105-133.
Bouanani B. Rekombinant DNA Teknolojisi ve Gelecekteki Bilim: Derleme makalesi/ Recombinant DNA technology and future science.:Vita Dergisi 2021 Sayı 1 S: 82-88.
Ali A, Kemter E, Wolf E. Advances in Organ and Tissue Xenotransplantation. Annu Rev Anim Biosci. 2024;15(12):369-390. doi: 10.1146/annurev-animal-021122-102606
Cassani G. Influence of the cellular host-vector system on the quality of therapeutic proteins obtained by recombinant DNA technology. Arzneimittelforschung. 1988; 38(5):762-4. PMID: 3415722.
Lindley SR, Subbaiah KCV, Priyanka F, et al. Ribozyme-activated mRNA trans-ligation enables large gene delivery to treat muscular dystrophies. Science. 2024; 386(6723):762-767. doi: 10.1126/science.adp8179.
Sayed N, Allawadhi P, Khurana A, et al. Gene therapy: Comprehensive overview and therapeutic applications. Life Sci. 2022; 1;294:120375. doi: 10.1016/j.lfs.2022.120375.
Terpe K. Overview of bacterial expression systems for heterologous protein production: from molecular and biochemical fundamentals to commercial systems. Appl Microbiol Biotechnol. 2006 Sep;72(2):211-22. doi: 10.1007/s00253-006-0465-8. Epub 2006 Jun 22. PMID: 16791589.
Schwartz MW, Seeley RJ, Campfield LA, et al. Identification of targets of leptin action in rat hypothalamus. J Clin Invest. 1996; 1;98(5):1101-6. doi: 10.1172/JCI118891.
Demain AL, Sanchez S. Microbial drug discovery: 80 years of progress. J Antibiot 2009;62(1):5-16. doi: 10.1038/ja.2008.16.
Rosano GL, Ceccarelli EA. Recombinant protein expression in Escherichia coli: advances and challenges. Front Microbiol. 2014; 17(5):172. doi: 10.3389/fmicb.2014.00172.
Fischer, S. The Role of Macroeconomic Factors in Growth. Journal of Monetary Economics, 1993; 32:485-512. http://dx.doi.org/10.1016/0304-3932(93)90027-d.
Jenkins N, Curling EM. Glycosylation of recombinant proteins: problems and prospects. Enzyme Microb Technol. 1994;16(5):354-64. doi: 10.1016/0141-0229(94)90149-x.
Diane MR, Jin, H. Lawrence Chew. Reliable protein production in a Pseudomonas fluorescens expression system. Protein Expression and Purification. 2012; 81(2), 157-165.
Anné J, Maldonado B, Van Impe J, et al.. Recombinant protein production and streptomycetes. J Biotechnol. 2012; 30:158(4):159-67. doi: 10.1016/j.jbiotec.2011.06.028.
Cereghino GP, Stark CM, Kim D, et al. The effect of α-mating factor secretion signal mutations on recombinant protein expression in Pichia pastoris. Gene. 2013; 519(2):311-7. doi: 10.1016/j.gene.2013.01.062.
Wu S, Letchworth GJ. High efficiency transformation by electroporation of Pichia pastoris pretreated with lithium acetate and dithiothreitol. Biotechniques. 2004;36(1):152-4. doi: 10.2144/04361DD02. PMID: 14740498.
Rajamanickam V, Metzger K, Schmid C, et al. A novel bi-directional promoter system allows tunable recombinant protein production in Pichia pastoris. Microb Cell Fact. 2017 :16(1):152. doi: 10.1186/s12934-017-0768-8.
Cos O, Ramón R, Montesinos J L, et al. Operational strategies, monitoring and control of heterologous protein production in the methylotrophic yeast Pichia pastoris under different promoters: a review. Microb Cell Fact. 2006; 6(5):17. doi: 10.1186/1475-2859-5-17.
Nocon J, Steiger M, Mairinger T, et al. Increasing pentose phosphate pathway flux enhances recombinant protein production in Pichia pastoris. Appl Microbiol Biotechnol. 2016;100(13):5955-63. doi: 10.1007/s00253-016-7363-5.
Kurtzman CP. Chapter 37 - Komagataella Y. Yamada, Matsuda, Maeda & Mikata (1995). The Yeasts (Fifth Edition) 2011, P 491-495.
Higgins DR, Cregg JM. Introduction to Pichia pastoris. Methods Mol Biol. 1998;103:1-15. doi: 10.1385/0-89603-421-6:1.
Mattanovich D, Branduardi P, Dato L, et al. Recombinant protein production in yeasts. Methods Mol Biol. 2012;824:329-58. doi: 10.1007/978-1-61779-433-9_17.
Prabhu AA, Veeranki VD. Metabolic engineering of Pichia pastoris GS115 for enhanced pentose phosphate pathway (PPP) flux toward recombinant human interferon gamma (hIFN-γ) production. Mol Biol Rep. 2018;45(5):961-972. doi: 10.1007/s11033-018-4244-2.
Macauley-Patrick S, Fazenda ML, McNeil B, et al. Heterologous protein production using the Pichia pastoris expression system. Yeast. 2005;22(4):249-70. doi: 10.1002/yea.1208.
Irina BS, James O, Olivia W. Rossanese, et al. A versatile set of vectors for constitutive and regulated gene expression in Pichia pastoris. Yeast. 1998;4(8) 15 June 1998 Pages 783-790.
Celik E and Calık P. Production of recombinant proteins by yeast cells. Biotechnol Adv, 2012;30(5):1108-18.
Looser V, Bruhlmann B, Bumbak F, et al . Cultivation strategies to enhance productivity of Pichia pastoris: A review. Biotechnology Advances 2015;33(6), Part 2, Pages 1177-1193.
Choi BK, Bobrowicz P, Davidson RC, et al. Use of combinatorial genetic libraries to humanize N-linked glycosylation in the yeast Pichia pastoris. Proc Natl Acad Sci. 2003,29;100(9):5022-7. doi: 10.1073/pnas.0931263100.
Nevalainen A, Seuri M. Of microbes and men. Indoor Air. 2005;15(9):58-64. doi: 10.1111/j.1600-0668.2005.00344.x.
Jenkins, JM., Astington, JW. Cognitive factors and family structure associated with theory of mind development in young children. Developmental Psychology, 1996;32(1), 70–78. https://doi.org/10.1037/0012-1649.32.1.70.
Meldgaard M, Svendsen I. Different effects of N-glycosylation on the thermostability of highly homologous bacterial (1,3-1,4)-beta-glucanases secreted from yeast. Microbiology (Reading). 1994;140(1):159-66. doi: 10.1099/13500872-140-1-159.
Gellissen G, Zbigniew A, Weydemann U, et al. High-level expression of foreign genes in Hansenula polymorpha. Biotechnology Advances, 1992; 10(2), 1992, Pages 179-189.
Wang Y, Liang ZH, Zhang YZ, et al Human insulin from a precursor overexpressed in the methylotrophic yeast Pichia pastoris and a simple procedure for purifying the expression product. Biotechnology and Bioengineering. 2001; 73(1) Pages 74-79.
Werten MW, Bosch TJ, Wind RD, et al. High-yield secretion of recombinant gelatins by Pichia pastoris. Yeast. 1999;15(11):1087-96. doi: 10.1002/(SICI)1097-0061(199908)15:11.
EasySelect™ Pichia Expression Kit For Expression of Recombinant Proteins Using pPICZ and pPICZα in Pichia pastoris Cat. no. K1740-01.
Kollabathula P, Katru S, Tumarada P, et al.. Harnessing Recombinant DNA Technology for Modern Innovations: A Review. International Journal of Dental Materials, 2025; 07, pp.27 - 31. 10.37983/ijdm.2025.7105.
Hocamurat H. Rekombinant DNA Teknolojisi ile Üretilen Peptid-Protein İlaçlar. Fabad J. Pharm. Sci. 1997; 22, 27-38.
Savaş HB , Fatih Gültekin F. İnsülin Direnci ve Klinik Önemi. Med J.2017:24(3):116-125 DOI:10.17343/sdutfd.264358.
Pfeifer GP, Singer-Sam J, Itakura K. et al. Nat Biotechnol. 2022;40(9):1317-1318. doi: 10.1038/s41587-022-01453-5.
Rabbani G, Ahn, SA. Structure, enzymatic activities, glycation and therapeutic potential of human serum albumin: a natural cargo. Int J Biol Macromol, 2019;123:979–990.
Cai K, Gierman, TM, Hotta J, et al. Ensuring the biologic safety of plasmaderived therapeutic proteins. BioDrugs 2005;19:79–96.
MacLennan S and Barbara. JA. Risks and side effects of therapy with plasma and plasma fractions. Best Pract. 2006;19,169-189.
Merten OW. Virus contaminations of cell cultures — a biotechnological view. Cytotechnology, 2002;39, 91-116.
Dudeja C, Mishra A, Ali A, et al. Microbial Genome Editing with CRISPR–Cas9: Recent Advances and Emerging Applications Across Sectors. Fermentation. 2025; 11(7):410. https://doi.org/10.3390/fermentation11070410.
Hektor HJ, Scholtmeijer K. Hydrophobins: proteins with potential. Curr Opin Biotechnol. 2005 Aug;16(4):434-9. doi: 10.1016/j.copbio.2005.05.004.
Wessels J, De Vries O, Asgeirsdottir S A, et al. Hydrophobin Genes Involved in Formation of Aerial Hyphae and Fruit Bodies in Schizophyllum. Plant Cell. 1991;3(8):793-799. doi: 10.1105/tpc.3.8.793.
Linder A, Christensson B, Herwarld H, et al. Protein: An Early Marker of Circulatory Failure in Sepsis. Clinical Infectious Diseases 2005, 49(7):1044-50 DOI:10.1086/605563.
Wösten HA. Hydrophobins: multipurpose proteins. Annu Rev Microbiol. 2001;55:625-46. doi: 10.1146/annurev.micro.55.1.625.
Praschak-Rieder N, Willeit M, Neumeister A et al. Prevalence of premenstrual dysphoric disorder in female patients with seasonal affective disorder. J Affect Disord. 2001;63(1-3):239-42. doi: 10.1016/s0165-0327(00)00176-2.
Sun Q. The Hydrophobic Effects: Our Current Understanding. Molecules. 2022; 18;27(20):7009. doi: 10.3390/molecules27207009.
Janssen I, Heymsfield SB, Wang ZM, et al. Skeletal muscle mass and distribution in 468 men and women aged 18-88 yr. J Appl Physiol. 2000;89(1):81-8. doi: 10.1152/jappl.2000.89.1.81.
Hanna JH, Saha K, Jaenisch R. Pluripotency and cellular reprogramming: facts, hypotheses, unresolved issues. Cell. 2010;12;143(4):508-25. doi: 10.1016/j.cell.2010.10.008.
Bayry J, Aimanianda V, Guijarro J I et al. Hydrophobins--unique fungal proteins. PLoS Pathog. 2012;8(5):e1002700. doi: 10.1371/journal.ppat.1002700.
Khalesi M, Gebruers K, Derdelinckx G. Recent Advances in Fungal Hydrophobin Towards Using in Industry. Protein J. 2015;34(4):243-55. doi: 10.1007/s10930-015-9621-2.
Kulkarni S, Nene S, Kalpana S. et al. Production of Hydrophobins from fungi. Process Biochemistry 61 DOI:10.1016/j.procbio.2017.06.012.
Ratcliffe NA, Furtado P J P, Dyson P, et al. Overview of paratransgenesis as a strategy to control pathogen transmission by insect vectors. Parasit Vectors. 2022;15(1):112. doi: 10.1186/s13071-021-05132-3.
Mirsalami SM, Mirsalami M, Advances in genetically engineered microorganisms: Transforming food production through precision fermentation and synthetic biology. Future Foods. 2025;11.
Strategies of strain improvement of industrial microbes: classical and recombinant DNA technology in improving the characteristics of industrially relevant microbes. Springer.
Advances in CRISPR-Cas technology and its applications. National Center for Biotechnology Information.
Advances in Therapeutic Applications of CRISPR Genome Editing .... National Center for Biotechnology Information.
Application of CRISPR-Cas9 genome editing technology in various .... National Center for Biotechnology Information.
Kursheed F, Naz E, Mateen S, et al. CRISPR applications in microbial World: Assessing the opportunities and challenges. Gene. 2025;935(30).
Matuszyńska A, Ebenhöh O, Matias D, et al. A new era of synthetic biology—microbial community design, Synthetic Biology, 2024; 9(1). https://doi.org/10.1093/synbio/ysae011.
Mariam I, Rova U, Christakopoulos P, et al. Data-driven synthetic microbes for sustainable future. NPJ Syst Biol Appl. 2025;11(1):74. doi: 10.1038/s41540-025-00556-4.
Scheffler R W. Asilomar Goes Underground: The Long Legacy of Recombinant DNA Hazard Debates for the Greater Boston Area Biotechnology Industry. J Hist Biol. 2025;58(1):67-93. doi: 10.1007/s10739-025-09806-x.
