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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 (rPA), has been designated as a significant breakthrough. Reteplase, 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 | ||
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