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Production of activ...
Production of active Exendin-4 in Nicotiana benthamiana and its application in treatment of type-2 diabetics
- Artikel/kapitelEngelska2022
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2022-12-22
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Frontiers Media SA,2022
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LIBRIS-ID:oai:prod.swepub.kib.ki.se:151515932
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http://kipublications.ki.se/Default.aspx?queryparsed=id:151515932URI
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https://doi.org/10.3389/fpls.2022.1062658DOI
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Språk:engelska
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Sammanfattning på:engelska
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GLP-1 (Glucagon-like peptide-1) is a peptide that stimulates insulin secretion from the β-cell for glycemic control of the plasma blood glucose level. Its mimetic exenatide (synthetic Exendin-4) with a longer half-life of approximately 3.3–4 h is widely used in clinical application to treat diabetes. Currently, exenatide is chemically synthesized. In this study, we report that the GLP-1 analogue recombinant Exendin-4 (Exdn-4) can be produced at a high level in Nicotiana benthamiana, with an estimated yield of 50.0 µg/g fresh biomass. For high-level expression, we generated a recombinant gene, B:GB1:ddCBD1m:8xHis : Exendin-4 (BGC : Exdn-4), for the production of Exendin-4 using various domains such as the BiP signal peptide, the GB1 domain (B1 domain of streptococcal G protein), a double cellulose binding domain 1 (CBD1), and 8 His residues (8xHis) to the N-terminus of Exendin-4. GB1 was used to increase the expression, whereas double CBD1 and 8xHis were included as affinity tags for easy purification using MCC beads and Ni2+-NTA resin, respectively. BGC : Exdn-4 was purified by single-step purification to near homogeneity using both Ni2+-NTA resin and microcrystalline cellulose (MCC) beads. Moreover, Exdn-4 without any extra residues was produced from BGC : Exdn-4 bound onto MCC beads by treating with enterokinase. Plant-produced Exdn-4 (Exendin-4) was as effective as chemically synthesized Exendin-4 in glucose-induced insulin secretion (GIIS) from mouse MIN6m9 cells a pancreatic beta cell line.
Biuppslag (personer, institutioner, konferenser, titlar ...)
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Afrin, S
(författare)
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Kim, J
(författare)
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Kang, JHY
(författare)
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Razzak, MA
(författare)
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Berggren, POKarolinska Institutet
(författare)
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Hwang, I
(författare)
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Karolinska Institutet
(creator_code:org_t)
Sammanhörande titlar
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Ingår i:Frontiers in plant science: Frontiers Media SA13, s. 1062658-1664-462X
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