Cover Image

Biologically active recombinant human erythropoietin expressed in hairy root cultures and regenerated plantlets of Nicotiana tabacum L.

Hairy root culture is a potential alternative to conventional mammalian cell culture to produce recombinant proteins due to its ease in protein recovery, low costs and absence of potentially human pathogenic contaminants. The current study focussed to develop a new platform of a hairy root culture s...

Full description

Saved in:
Bibliographic Details
Main Authors: Gurusamy, Poornima Devi (Author) , Schäfer, Holger (Author) , Wink, Michael (Author)
Format: Article (Journal)
Language:English
Published: August 11, 2017
In: PLOS ONE
Year: 2017, Volume: 12, Issue: 8
ISSN:1932-6203
DOI:10.1371/journal.pone.0182367
Online Access:Verlag, kostenfrei, Volltext: http://dx.doi.org/10.1371/journal.pone.0182367
Verlag, kostenfrei, Volltext: http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0182367
Get full text
Author Notes:Poornima Devi Gurusamy, Holger Schäfer, Siva Ramamoorthy, Michael Wink

MARC

LEADER 00000caa a2200000 c 4500
001 1565915364
003 DE-627
005 20230426064405.0
007 cr uuu---uuuuu
008 171130s2017 xx |||||o 00| ||eng c
024 7 |a 10.1371/journal.pone.0182367  |2 doi 
035 |a (DE-627)1565915364 
035 |a (DE-576)49591536X 
035 |a (DE-599)BSZ49591536X 
035 |a (OCoLC)1263948952 
040 |a DE-627  |b ger  |c DE-627  |e rda 
041 |a eng 
084 |a 32  |2 sdnb 
100 1 |a Gurusamy, Poornima Devi  |e VerfasserIn  |0 (DE-588)1139193848  |0 (DE-627)896838137  |0 (DE-576)492984034  |4 aut 
245 1 0 |a Biologically active recombinant human erythropoietin expressed in hairy root cultures and regenerated plantlets of Nicotiana tabacum L.  |c Poornima Devi Gurusamy, Holger Schäfer, Siva Ramamoorthy, Michael Wink 
264 1 |c August 11, 2017 
300 |a 23 
336 |a Text  |b txt  |2 rdacontent 
337 |a Computermedien  |b c  |2 rdamedia 
338 |a Online-Ressource  |b cr  |2 rdacarrier 
500 |a Gesehen am 30.11.2017 
520 |a Hairy root culture is a potential alternative to conventional mammalian cell culture to produce recombinant proteins due to its ease in protein recovery, low costs and absence of potentially human pathogenic contaminants. The current study focussed to develop a new platform of a hairy root culture system from Nicotiana tabacum for the production of recombinant human EPO (rhEPO), which is regularly produced in mammalian cells. The human EPO construct was amplified with C-terminal hexahistidine tag from a cDNA of Caco-2 cells. Two versions of rhEPO clones, with or without the N-terminal calreticulin (cal) fusion sequence, were produced by cloning the amplified construct into gateway binary vector pK7WG2D. Following Agrobacterium rhizogenes mediated transformation of tobacco explants; integration and expression of constructs in hairy roots were confirmed by several tests at DNA, RNA and protein levels. The amount of intracellular rhEPO from hairy root cultures with cal signal peptide was measured up to 66.75 ng g-1 of total soluble protein. The presence of the ER signal peptide (cal) was essential for the secretion of rhEPO into the spent medium; no protein was detected from hairy root cultures without ER signal peptide. The addition of polyvinylpyrrolidone enhanced the stabilization of secreted rhEPO leading to a 5.6 fold increase to a maximum concentration of 185.48 pg rhEPOHR g-1 FW hairy root cultures. The rhizo-secreted rhEPO was separated by HPLC and its biological activity was confirmed by testing distinct parameters for proliferation and survival in retinal pigment epithelial cells (ARPE). In addition, the rhEPO was detected to an amount 14.8 ng g-1 of total soluble leaf protein in transgenic T0 generation plantlets regenerated from hairy root cultures with cal signal peptide. 
650 4 |a Cloning 
650 4 |a Erythropoietin 
650 4 |a Genetically modified plants 
650 4 |a Hair 
650 4 |a Leaves 
650 4 |a Nicotiana 
650 4 |a Recombinant proteins 
650 4 |a Signal peptides 
700 1 |a Schäfer, Holger  |d 1969-  |e VerfasserIn  |0 (DE-588)1067525173  |0 (DE-627)818856912  |0 (DE-576)426714911  |4 aut 
700 1 |a Wink, Michael  |d 1951-  |e VerfasserIn  |0 (DE-588)124618146  |0 (DE-627)363448519  |0 (DE-576)164659390  |4 aut 
773 0 8 |i Enthalten in  |t PLOS ONE  |d San Francisco, California, US : PLOS, 2006  |g 12(2017,8) Artikel-Nummer e0182367, 23 Seiten  |h Online-Ressource  |w (DE-627)523574592  |w (DE-600)2267670-3  |w (DE-576)281331979  |x 1932-6203  |7 nnas  |a Biologically active recombinant human erythropoietin expressed in hairy root cultures and regenerated plantlets of Nicotiana tabacum L. 
773 1 8 |g volume:12  |g year:2017  |g number:8  |g extent:23  |a Biologically active recombinant human erythropoietin expressed in hairy root cultures and regenerated plantlets of Nicotiana tabacum L. 
856 4 0 |u http://dx.doi.org/10.1371/journal.pone.0182367  |x Verlag  |x Resolving-System  |z kostenfrei  |3 Volltext 
856 4 0 |u http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0182367  |x Verlag  |z kostenfrei  |3 Volltext 
951 |a AR 
992 |a 20171130 
993 |a Article 
994 |a 2017 
998 |g 124618146  |a Wink, Michael  |m 124618146:Wink, Michael  |d 160000  |d 160100  |d 700000  |d 729400  |e 160000PW124618146  |e 160100PW124618146  |e 700000PW124618146  |e 729400PW124618146  |k 0/160000/  |k 1/160000/160100/  |k 0/700000/  |k 1/700000/729400/  |p 4  |y j 
998 |g 1067525173  |a Schäfer, Holger  |m 1067525173:Schäfer, Holger  |d 160000  |d 160100  |e 160000PS1067525173  |e 160100PS1067525173  |k 0/160000/  |k 1/160000/160100/  |p 2 
998 |g 1139193848  |a Gurusamy, Poornima Devi  |m 1139193848:Gurusamy, Poornima Devi  |d 160000  |d 160100  |e 160000PG1139193848  |e 160100PG1139193848  |k 0/160000/  |k 1/160000/160100/  |p 1  |x j 
999 |a KXP-PPN1565915364  |e 298853652X 
BIB |a Y 
SER |a journal 
JSO |a {"physDesc":[{"extent":"23 S."}],"recId":"1565915364","origin":[{"dateIssuedDisp":"August 11, 2017","dateIssuedKey":"2017"}],"relHost":[{"physDesc":[{"extent":"Online-Ressource"}],"recId":"523574592","disp":"Biologically active recombinant human erythropoietin expressed in hairy root cultures and regenerated plantlets of Nicotiana tabacum L.PLOS ONE","origin":[{"dateIssuedKey":"2006","dateIssuedDisp":"2006-","publisherPlace":"San Francisco, California, US ; Lawrence, Kan.","publisher":"PLOS ; PLoS"}],"type":{"media":"Online-Ressource","bibl":"periodical"},"language":["eng"],"note":["Schreibweise des Titels bis 2012: PLoS ONE","Gesehen am 20.03.19"],"title":[{"title_sort":"PLOS ONE","title":"PLOS ONE"}],"corporate":[{"display":"Public Library of Science","role":"isb"}],"name":{"displayForm":["Public Library of Science"]},"pubHistory":["1.2006 -"],"id":{"eki":["523574592"],"issn":["1932-6203"],"zdb":["2267670-3"]},"part":{"issue":"8","extent":"23","volume":"12","text":"12(2017,8) Artikel-Nummer e0182367, 23 Seiten","year":"2017"}}],"person":[{"display":"Gurusamy, Poornima Devi","role":"aut","given":"Poornima Devi","family":"Gurusamy"},{"display":"Schäfer, Holger","family":"Schäfer","given":"Holger","role":"aut"},{"display":"Wink, Michael","family":"Wink","role":"aut","given":"Michael"}],"type":{"media":"Online-Ressource","bibl":"article-journal"},"language":["eng"],"note":["Gesehen am 30.11.2017"],"title":[{"title_sort":"Biologically active recombinant human erythropoietin expressed in hairy root cultures and regenerated plantlets of Nicotiana tabacum L.","title":"Biologically active recombinant human erythropoietin expressed in hairy root cultures and regenerated plantlets of Nicotiana tabacum L."}],"name":{"displayForm":["Poornima Devi Gurusamy, Holger Schäfer, Siva Ramamoorthy, Michael Wink"]},"id":{"doi":["10.1371/journal.pone.0182367"],"eki":["1565915364"]}} 
SRT |a GURUSAMYPOBIOLOGICAL1120 

Similar Items