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Journals / Türk Biyokimya Dergisi / 2019 / Cilt: 44 - Sayı: 3

Expression of a functional recombinant vascular endothelial growth factor 165 (VEGF165) in Arabidopsis thaliana

Arabidopsis thaliana’daki fonksiyonel bir rekombinant vasküler endotelyal büyüme faktörü 165’in (VEGF165) ekspresyonu

Xi CHEN
Xinxin LAN
Kun JI
Jing YANG
Haiyan LI
Xiaoyu SUN
Xue FENG
Linna DU
Weidong QIANGA
Pages
254–260
Publication Date
2022-11-04
DOI
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AI Summary & Key Takeaways

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⚡ TL;DR(Too Long; Didn't Read)

Finally, MTT assays showed that the oleosin-VEGF165 fusion protein played a part in the proliferation of HUVEC cells in vitro.Conclusion: Oleosin-VEGF165 was successfully expressed and it had stimulated HUVEC cell proliferation activity.

📌 3 Key Takeaways

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Objective & Hypothesis:Objective: Targeting the protein of interest to a particular tissue to achieve high-level expression is an important strategy to increase expression efficiency.
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Key Findings & Data:The use of the plant seed oil body as a bioreactor can not only increase the amount of target protein, but also reduce the cost of downstream processing.Methods: VEGF165 was expressed in Arabidopsis thalianaseeds via oilbody fusion technology.
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Conclusion & Impact:Finally, MTT assays showed that the oleosin-VEGF165 fusion protein played a part in the proliferation of HUVEC cells in vitro.Conclusion: Oleosin-VEGF165 was successfully expressed and it had stimulated HUVEC cell proliferation activity.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:Domain Parameters

Abstract

Objective: Targeting the protein of interest to a particular tissue to achieve high-level expression is an important strategy to increase expression efficiency. The use of the plant seed oil body as a bioreactor can not only increase the amount of target protein, but also reduce the cost of downstream processing.Methods: VEGF165 was expressed in Arabidopsis thalianaseeds via oilbody fusion technology. The pKO-VEGF165vector was construted and transformed into A. thalianaseeds. T3 transgenic seeds was detected by SDS-PAGE and western blot methods. The cell activity was tested by MTT methods.Result: The phaseolin promoter was used to drive seed-specific expression of the VEGF165 gene in transgenic A. thaliana. The coding region of VEGF165 was fused to the Arabidopsis oleosin sequence to target the protein to the oil bodies in the seeds of transgenic plants. The T-DNA region of recombinant plasmid pKO-VEGF165 was shifted to A. thaliana seeds via the floral-dip method. Protein was analyzed by electrophoresis and protein hybridization analyses. Finally, MTT assays showed that the oleosin-VEGF165 fusion protein played a part in the proliferation of HUVEC cells in vitro.Conclusion: Oleosin-VEGF165 was successfully expressed and it had stimulated HUVEC cell proliferation activity.

Özet

Amaç: Yüksek seviyeli ekspresyon elde etmek için ilgili proteininbelirlibirdokuyahedeflenmesi,ekspresyonverimliliğini artırmak için önemli bir stratejidir. Bir biyo-reaktör olarak bitki tohum yağı gövdesinin kullanılması, sadece hedef protein miktarını arttırmakla kalmaz, aynı zamanda aşağı akış işleme maliyetini de düşürür.Metod: VEGF165, yağlı vücut füzyon teknolojisi vasıta-sıyla Arabidopsis thaliana tohumlarında eksprese edildi. pKO-VEGF165 vektörü dizilmiş ve Arabidopsis thalianatohumlarına dönüştürülmüştür. T3 transjenik tohumları SDS-PAGE ve western blot yöntemleri ile tespit edildi. Hücre aktivitesi MTT yöntemleriyle test edildi. Protein, elektroforez ve protein hibridizasyon analizleri ile analiz edildi Bulgular: Transazensel Arabidopsis thaliana’daki tohum-dan spesifik ekspresyonunu başlatmak için fazoin yük-seltici kullanılmıştır. VEGF165, in kodlama bölgesi, prote-ini transjenik bitkilerin tohumlarındaki yağ gövdelerine hedeflemekiçinArabidopsisoleosindizisinekaynaş-tırıldı.RekombinantplazmidpKO-VEGF165,inT-DNAbölgesi, çiçek-daldırma yöntemi vasıtasıyla Arabidopsis thaliana tohumlarına kaydırılmıştır. Finally, MTT assays showed that the oleosin-VEGF165 fusion protein played a part in the proliferation of HUVEC cells in vitro.Sonuç:Oleozin-VEGF165başarıylaekspreseedildiveHUVEC hücre çoğalması aktivitesini uyardı.

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Cite This Article

CHEN, X., LAN, X., JI, K., YANG, J., LI, H., SUN, X., FENG, X., DU, L., QIANGA, W. (2022). Expression of a functional recombinant vascular endothelial growth factor 165 (VEGF165) in Arabidopsis thaliana. Türk Biyokimya Dergisi, Cilt: 44 - Sayı: 3, 254–260. https://ajindex.com/articles/620835

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