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Titre du document / Document title

The stability of pET21 + PA, a pET derived plasmid, under different culture conditions

Auteur(s) / Author(s)

Vahedi Fatemeh (1) ; Khatami Fatemeh (1) ; Es-haghi Ali (1) ;

Affiliation(s) du ou des auteurs / Author(s) Affiliation(s)

(1) Razi Vaccine & Serum Research Institute, Mashhad, Iran

Résumé / Abstract

The recombinant plasmid pET21 + PA that has been deposited at Genbank with accession number EF550209 was constructed by inserting the 1,700-bp PA (protective antigen of Bacillus anthracis) recombinant gene into Xho I/Hind Ш sites of the pET21b + vector under the control of the T7 promoter for highly expressing PA. pET21 + PA was cloned into Escherichia coli BL21 strain. The high activity of T7 RNA polymerase could make a powerful expression system for high-level expression of the recombinant proteins. However, during the large-scale production of recombinant proteins, the productivity of a fermentation process is directly affected by many factors, such as plasmid stability, protein production, and culture conditions. In this study, we studied the effects of various culture conditions on the plasmid stability and target protein yield including antibiotic concentrations, the time of induction by IPTG, and the number of successive cultures. The results indicated that the plasmid pET21 + PA is completely stable after the fiftieth generation. Loss of plasmid and structural change were not detected but the yield of protein production was decreased by about 10% in generation 50. These data would be useful for the industrial production of the recombinant PA vaccine and other recombinant proteins.

Revue / Journal Title

World journal of microbiology & biotechnology    ISSN  1573-0972 

Source / Source

2011, vol. 27, no2, pp. 253-257 [5 page(s) (article)]

Langue / Language

Anglais

Editeur / Publisher

Springer, Heidelberg, ALLEMAGNE  (2002) (Revue)

Mots-clés d'auteur / Author Keywords

Plasmid stability

;

High-level expression

;

Protective antigen

;

E. coli

;

Localisation / Location

35400060672640.0009

Springer Science+Business Media B.V., 2011
Nº notice refdoc (ud4) : 23597026



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