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Transient expression of reteplase gene (rPA) under methyl jasmonat treatment in cucumber (Cucumis sativus L.) | ||
| Iranian Journal of Genetics and Plant Breeding | ||
| دوره 12، شماره 1 - شماره پیاپی 23، تیر 2023، صفحه 77-85 اصل مقاله (650.42 K) | ||
| نوع مقاله: Research paper | ||
| شناسه دیجیتال (DOI): 10.30479/ijgpb.2023.18604.1337 | ||
| نویسندگان | ||
| Maryam Ghayomi Abkenar* 1؛ Mokhtar Jalali Javaran1؛ Hamideh Ofoghi2 | ||
| 1Department of Plant Breeding and Biotechnology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran. | ||
| 2Department of Biotechnology, Iranian Research Organization for Science and Technology, Tehran, Iran. | ||
| تاریخ دریافت: 22 فروردین 1402، تاریخ بازنگری: 24 شهریور 1402، تاریخ پذیرش: 05 مهر 1402 | ||
| چکیده | ||
| Every year, millions of people are suffering from cardiovascular diseases. Thrombolytic and fibrinolytic therapies have shown results in the treatment of cardiovascular diseases and strokes. Tissue plasminogen activator (tPA) is a pivotal thrombolytic drug that plays a crucial role in cardiovascular disorders. The recombinant version of tPA, known as alteplase, has been designated as a significant breakthrough. Reteplase (rPA) a third-generation mutant of alteplase, has also emerged as a notable variant. The objective of the present study was to design a transient expression system to analyze the gene expression of rPA in cucumber plants. Moreover, the effect of elicitor methyl jasmonate (MeJA) was assessed on rPA gene expression. To do this, the rPA gene was amplified and subcloned into the ZYMV vector. The recombinant vector was then inoculated in cucumber leaves, and the transfected leaves were harvested at three, five, and ten days post-inoculation (DPI). An RT-PCR reaction was performed, and the desired band was observed in the inoculated lines. Furthermore, the quantity of rPA protein expression was assessed using ELISA. The results of the study demonstrated that the presence of methyl jasmonate led to an enhancement in the production of recombinant protein in the inoculated samples. The highest level of rPA gene expression was observed in the samples harvested on the tenth day post-inoculation with methyl jasmonate treatment which was 450.83 ng µg-1 of total plant-extracted protein. For the first time, the rPA protein was successfully expressed under the influence of the methyl jasmonate elicitor in the cucumber plants. | ||
| کلیدواژه‌ها | ||
| Gene expression؛ Methyl jasmonate Elicitor؛ Pharming؛ Recombinant protein؛ Reteplase؛ Transient expression | ||
| عنوان مقاله [English] | ||
| بیان موقت ژن رتپلاز (rPA) تحت القاگر متیل جاسمونات در گیاه خیار (Cucumis Sativus L.) | ||
| نویسندگان [English] | ||
| مریم قیومی آبکنار1؛ مختار جلالی جواران1؛ حمیده افقی2 | ||
| 1گروه اصلاح نباتات و بیوتکنولوژی، دانشکده کشاورزی، دانشگاه تربیت مدرس، تهران، ایران. | ||
| 2گروه بیوتکنولوژی، سازمان تحقیقات علم و فناوری ایران، تهران، ایران. | ||
| چکیده [English] | ||
| هرساله میلیون ها نفر از بیماریهای قلبی-عروقی رنج میبرند. درمان های ترومبولیتیک و فیبرینولیتیک نتایجی را در درمان بیماری های قلبی عروقی نشان داده اند. فعال کننده پلاسمینوژن بافتی (tPA) یک داروی ترومبولیتیک حیاتی است که نقش مهمی در رسیدگی به اختلالات قلبی عروقی دارد. tPA نوترکیب با عنوان آلتپلاز شناخته شده است. رتپلاز (rPA) نسل سوم جهش یافته آلتپلاز، به عنوان یک واریانت بسیار مهم می‌باشد. هدف از مطالعه حاضر، طراحی سیستم بیان موقت برای انالیز بیان ژن rPA در گیاهان خیار است و الیسیتور متیل جاسمونات برای ارزیابی تأثیر آن در بیان ژن، مورد ازمایش قرار گرفت. برای روش تحقیق، ژن rPA تکثیر و در وکتور ZYMV کلون شد. وکتور نوترکیب ایحاد شده به برگ های گیاه خیار تلقیح شد. و برگ های تلقیح شده در سه، پنج و ده روز پس از تلقیح (DPI) برداشت شدند. علاوه بر این واکنش RT-PCR انجام شد و باند مورد نظر در لاین های تلقیح شده مشاهده شد. و کیفیت پروتئین های رتپلاز بیان شده با ازمایش الایزا مورد بررسی قرار گرفت. نتایج این مطالعه نشان داد که حضور متیل جاسمونات منجر به افزایش تولید پروتئین نوترکیب در نمونه‌های تلقیح شده میشود. قابل ذکر است که بالاترین سطح بیان ژن rPA در نمونه های برداشت شده در روز دهم پس از تلقیح (DPI) و تیمار شده با متیل جاسمونات مشاهده شد که مقدار ان 450.83 ng/µg از کل پروتئین های استخراج شده گیاه بود. برای اولین بار، پروتئین rPA تحت تأثیر الیسیتور متیل جاسمونات با موفقیت در گیاه خیار بیان شد. | ||
| کلیدواژه‌ها [English] | ||
| بیان موقت ژن, الیسیتور متیل جاسمونات, فارمینگ, پروتئین نوترکیب, رتپلاز, بیان موقت | ||
| مراجع | ||
|
Benfey P. N., and Chua N. H. (1990). The cauliflower mosaic virus 35s promoter: combinatorial regulation of transcription in plants. Science, 250(4983): 959-66. Bradford M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72: 248-254. Canto T. (2016). Transient expression systems in plants: potentialities and constraints. Advances in Experimental Medicine and Biology, 896: 287-301. Clark D. P., and Pazdernik N. J. (2016). Biotechnology academic cell. Second Edition, USA: Washington University School of Medicine, Chapter 3, pp. 825. Daniell H., Singh N. D., Mason H., and Streatfield S. J. (2009). Plantmade vaccine antigens and biopharmaceuticals. Trends Plant Science, 14(12): 669- 679. Egelkrout E., Rajan V., and Howard J. A. (2012). Overproduction of recombinant proteins in plants. Plant Science, 184: 83-101. Engvall E., and Perlman P. (1971). Enzyme-linked immune sorbent assay (ELISA). Quantitative assay of immunoglobulin G. Immunochemistry, 8(9): 871-874. Feng Z., Li X., Fan B., Zhu C., and Chen Z. (2022). Miximizing the production of recombinant proteins in plants: from transcription to protein stability. Intrenational Journal of Molecular Sciences, 23(21): 13516. Fischer R., and Buyel J. F. (2020). Molecular farming-The slope of enlightenment. Biotechnology Advances, 40: 107519. Gengenbach B. B., Keil L. L., Opdensteinen P., Müschen C. R., Melmer G., Lentzen H., Bührmann J., and Buyel J. F. (2019). Comparison of microbial and transient expression for the production and purification of the anticancer mistletoe lectin viscumin. Biotechnol Bioeng, 116: 2236-2249. Gleba Y., Klimyuk V., and Marillonnet S. (2007). Viral vectors for the expression of proteins in plants. Current Opinion in Biotechnology, 18: 134-141. Gould K. (2022). Growing cucumbers: how to add crunch to your garden this season. Treehugger. https://www.treehugger.com/how-to-grow-cucumbers-5195302#toc-how-to-harvest-store-and-preserve-cucumbers. Guy C. L., and Haskell D. (1992). Detection of polypeptides associated with the cold acclimation process in spinach. Electrophoresis, 9(11): 787-796. Hirano S. (2012). Western blot analysis. Methods in Molecular Biology, 926: 87-97. Hsu C-H., Lin S-S., Liu F-L., Su W-C., and Yeh S-D. (2004). Oral administration of a mite allergen expressed by Zucchini yellow mosaic virus in cucurbit species downregulates allergen-induced airway inflammation and IgE synthesis. Journal of Allergy and Clinical Immunology, 113: 1079-1085. Huang S., Li R., Zhang Z., Li L., et al. (2009). The genome of the cucumber, Cucumis sativus L. Nature Genetics, 41: 1275-1283. Kanamoto H., Yamashita A., Asao H., Okumura S., Takase H., Hattori M., Yokota A., and Tomizawa K. (2006). Efficient and stable transformation of Lactuca Sative L. cv. cisco (lettuce) plastids. Transgenic Research, 15: 205-2177. Kelada K. D., Tusé D., Gleba Y., McDonald K. A., and Nandi S. (2021). Process simulation and techno-economic analysis of large scale bioproduction od sweet protein thaumatin. Foods, 10: 838. Kohnert U. (1992). Biochemical properties of the kringle2 and protease domains are maintained in the refolded tPA deletion variant BM 06.022. Protein Engineering, 5(1): 93-100. Lai E. M., Shih H. W., Wen S. R., Cheng M. W., Hwang H. H., and Chiu S. H. (2006). Proteomic analysis of Agrobacterium tumefaciens response to the Vir gene inducer acetosyringone. Proteomics, 6(14): 4130-6. Ma T., Li Z., and Wang S. (2019). Production of bioactive recombinant reteplase by virus-based transient expression system in Nicotiana benthamiana. Front Plant Science, 10: 1225. Makhzoum A., Benyammi R., Moustafa K., and Trémouillaux-Guiller J. (2014). Recent advances on host plants and expression cassettes’ structure and function in plant molecular pharming. BioDrugs, 28: 145-159. Moustafa K., Makhzoum A., and Trémouillaux-Guiller J. (2016). Molecular farming on rescue of pharma industry for next generation. Critical Reviews in Biotechnology, 36(5): 840-850. Nikitin D., Choi S., Mican J., Toul M., Ryu W. S., Damborsky J., Mikulik R., and Kim D. E. (2021). Development and testing of thrombolytics in stroke. Journal of Stroke, 23(1): 12-36. Nosaki S., Hoshikawa K., Ezura H., and Miura K. (2021). Transient protein expression systems in plants and their aplications. Plant Biotechnol (Tokyo), 38(3): 297-304. Parcq J., Bertrand T., Baron A. F., Hommet Y., Anglès-Cano E., and Vivien D. (2013). Molecular requirements for safer generation of thrombolytics by bioengineering the tissue-type plasminogen activator A chain. Journal of Thrombosis and Haemostasis, 11: 539-546. Robert S., Goulet M. C., D’Aoust M. A., Sainsbury F., and Michaud D. (2015). Leaf proteome rebalancing in Nicotiana benthamiana for upstream enrichment of a transiently expressed recombinant protein. Plant Biotechnology Journal, 13: 1169-1179. Rodriguez-Concepcion M., and Daròs J. A. (2022). Transient expression systems to rewire plant csrotenoid metabolism. Current Opinion in Plant Biology, 66: 102190. Roth M., and Manwani D. (2011). Thrombotic desorders. Pediatrics in Review, 32: 41-43. DOI: 10.1542/pir.32-1-41. Sablowski R. W., Baulcombe D. C., and Bevan M. (1995). Expression of a flower-specific Myb protein in leaf cells using a viral vector causes ectopic activation of a target promoter. Proceeding of the National Academy of Sciences of the United States of America, 92: 6901-6905. Sala F., Rigano M. M., Barbante A., Basso B., Walmsley A. M., and Castiglions S. (2003). Vaccine antigen production in transgenic plants: Strategies, gene construsts and perspectives. Vaccine, 21: 803-808. Sambrook J., and Russell D. W. (2001). Molecular cloning, a laboratory manual. 3rd Edition, Cold Spring Harbor Laboratory Press, New York, pp. 2344. Schillberg S., and Finnern R. (2021). Plant molecular farming for the production of valuable proteins - Critical evaluation of achievements and future challenges. Journal of Plant Physiology, 258-259: 153359. Schillberg S., and Spiegel H. (2022). Recombinant protein production in plants: a brief owerview of strengths and challenges. Methods in Molecular Biology, 2480: 1-13. Wang S., Ku S. S., YE X., HE C., Kwon S. Y., and Choi P. S. (2015). Current status of genetic transformation technology developed in cucumber (Cucumis Sativus L.). Journal of Integrative Agriculture, 14(3): 469-482. World Health Organization, https://www.who.int/health-topics/cardiovascular-diseases. Xu J., Dolan M. C., Medrano G., Cramer C. L., and Weathers P. J. (2012). Green factory: plants as bioproduction platforms for recombinant proteins. Biotechnology Advances, 30: 1171- 1184. Yamamoto T., Hoshikawa K., Ezura K., Okazawa R., et al. (2018). Improvement of the transient expression system for the production of recombinant proteins in plants. Scientific Reports, 8: 1-10. Zhang J., Feng S., Yuan J., Wang Ch., Lu T., Wang H., and Yu Ch. (2021). The formation of fruit quality in Cucumis Sativus L. Frontiers in Plant Science, 12: 729448. | ||
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