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Anti-VEGFR2 protein accumulation under the influence of methyl jasmonate treatment in Brassica napus and Nicotiana benthamiana plants | ||
| Iranian Journal of Genetics and Plant Breeding | ||
| دوره 11، شماره 2 - شماره پیاپی 22، دی 2022، صفحه 59-67 اصل مقاله (725.98 K) | ||
| نوع مقاله: Research paper | ||
| شناسه دیجیتال (DOI): 10.30479/ijgpb.2023.18804.1341 | ||
| نویسندگان | ||
| Mahsa Sadri1؛ Mokhtar Jalali Javaran* 1؛ Mojgan Soleimanizadeh2 | ||
| 1Department of Agriculture Biotechnology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran. | ||
| 2Department of Horticultural Science, Faculty of Agriculture and Natural Resources, University of Hormozgan, Bandar Abbas, Iran. | ||
| تاریخ دریافت: 08 خرداد 1402، تاریخ بازنگری: 03 مهر 1402، تاریخ پذیرش: 03 مهر 1402 | ||
| چکیده | ||
| This study was performed to investigate the effect of methyl jasmonate (MeJA) elicitor on green fluorescent protein (GFP) and Anti vascular endothelial growth factor (Anti-VEGFR2) proteins accumulation. A factorial experiment based on completely randomized design (CRD) was performed primarily to evaluate the effect of MeJA and days after inoculation on GFP expression in Brassica napus and Nicotiana benthamiana. RT-PCR and qRt-PCR assay was performed for GFP gene transcription confirmation at mRNA level. ELISA was done to investigate effect of the elicitor on GFP and Anti-VEGFR2 accumulation. The optimum treatment combination was then used to check Anti-VEGFR2 accumulation. GFP expression was significantly enhanced using M1D10 combination (the best treatment combination) in the two plants. This increase was greater in Brassica napus than in Nicotiana benthamiana. GFP and Anti-VEGFR2 transcript levels showed no significant differences between the optimized and control treatments in the qRT-PCR experiment, but the quantity of Anti-VEGFR2 protein accumulated in optimized treatments was higher than in control. A simple explanation for the improved accumulation of GFP and Anti-VEGFR2 would be a low ‘sink pressure’ on amino acid pools towards RuBisCO biosynthesis in RuBisCO-depleted leaves, and a resulting increased availability of metabolite and cellular resources for the production of less abundant (e.g., recombinant) proteins. This research showed that MeJA elicitor increased the accumulation of both proteins. | ||
| کلیدواژه‌ها | ||
| Elicitor؛ GFP؛ Molecular farming؛ Recombinant protein؛ Transient expression | ||
| عنوان مقاله [English] | ||
| تجمع پروتئین نوترکیب Anti-VEGFR2 تحت تاثیر تیمار متیل جاسمونات درگیاهان کلزا و توتون | ||
| نویسندگان [English] | ||
| مهسا صدری1؛ مختار جلالی جواران1؛ مژگان سلیمانی زاده2 | ||
| 1گروه بیوتکنولوژی کشاورزی، دانشکده کشاورزی، دانشگاه تربیت مدرس، تهران، ایران. | ||
| 2گروه علوم باغبانی، دانشکده کشاورزی، دانشگاه هرمزگان، بندرعباس، ایران. | ||
| چکیده [English] | ||
| این تحقیق به‌منظور بررسی اثر الیسیتور متیل جاسمونات (MeJA) روی تجمع پروتئین‌های نوترکیب GFP و Anti-VEGFR2 انجام شد. ابتدا به‌منظور ارزیابی اثر MeJA و روزهای نمونه‌برداری پس از مایه‌کوبی روی بیان GFP در کلزا و توتون آزمایش فاکتوریلی در قالب طرح کاملا تصادفی انجام شد. برای تأیید رونویسی ژن GFP در سطح RNA، RT-PCR و آزمون qRt-PCR انجام شد. آزمون الیزا برای بررسی اثر الیسیتور روی تجمع GFP و Anti-VEGFR2 انجام شد. به‌منظور تأیید بیان ژن GFP در سطح پروتئین و سپس تعیین بهترین ترکیب تیماری، بیان ژن GFP تحت تاثیر تمام ترکیبات با استفاده از الیزا بررسی شد. سپس بهترین ترکیب تیماری برای بررسی تجمع Anti-VEGFR2 استفاده شد. بیان GFP به‌طور معنی‌داری با استفاده از ترکیب M1D10 (بهترین ترکیب تیماری) در هر دو گیاه افزایش یافت. این افزایش در گیاه کلزا بیشتر از توتون بود. بنابراین بهترین ترکیب تیماری برای بیان ژن Anti-VEGFR2 در گیاهان کلزا استفاده شد. در آزمایش qRT-PCR سطوح رونوشت GFP و Anti-VEGFR2 هیچ گونه تفاوت معنی‌داری بین تیمارهای شاهد و بهینه نشان نداد، اما مقدار پروتئین Anti-VEGFR2 تجمع یافته در تیمارهای بهینه شده بیشتر از کنترل بود. توضیح ساده برای تجمع افزایش یافته نانوبادی Anti-VEGFR2 و GFP کاهش بیان پروتئین RuBisCO در برگ‌ها و در نتیجه افزایش دسترسی منابع متابولیکی و سلولی برای تولید پروتئین‌های با فراواتی کمتر (از جمله پروتئین‌های نوترکیب) میباشد. این تحقیق نشان داد که الیسیتور تجمع هر دو پروتئین نوترکیب GFP و Anti-VEGFR2 را افزایش می‌دهد. | ||
| کلیدواژه‌ها [English] | ||
| الیسیتور, بیان موقت, پروتئین نوترکیب, زراعت مولکولی | ||
| مراجع | ||
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