Secretion of hepatitis B surface antigen in transformed tobacco cell suspension cultures

Six different expression cassettes of hepatitis B surface antigen (HBsAg) were used to transform tobacco cell suspension cultures. The transgenic nature of the cells was confirmed by PCR. The secreted HBsAg was assayed by ELISA and analyzed by Western blotting. A maximum of 31 microg antigen/l was o...

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Bibliographic Details
Published in:Biotechnology letters 2005-07, Vol.27 (13), p.927-932
Main Authors: SUNIL KUMAR, G. B, GANAPATHI, T. R, SRINIVAS, L, REVATHI, C. J, BAPAT, V. A
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Blotting, Western
Cell Culture Techniques
Cell Line
Cyclopentanes - pharmacology
DNA, Plant - analysis
DNA, Plant - genetics
Fundamental and applied biological sciences. Psychology
Hepatitis
Hepatitis B Surface Antigens - genetics
Hepatitis B Surface Antigens - metabolism
Nicotiana - cytology
Nicotiana - genetics
Nicotiana - metabolism
Oxylipins
Peptides
Polymerase Chain Reaction
Recombinant Fusion Proteins - drug effects
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
Salicylic Acid - pharmacology
Transformation, Genetic
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Summary:Six different expression cassettes of hepatitis B surface antigen (HBsAg) were used to transform tobacco cell suspension cultures. The transgenic nature of the cells was confirmed by PCR. The secreted HBsAg was assayed by ELISA and analyzed by Western blotting. A maximum of 31 microg antigen/l was obtained in the spent medium from the transformed cells. The use of an ethylene-forming enzyme promoter and incorporation of C-terminal endoplasmic-reticulum-retention signal enhanced the secretion of HBsAg. Salicylic or jasmonic acid at 10 microM: increased secretion of HBsAg by six fold.
ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-005-7185-5