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 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.
Kumar
G B Sunil
GB
Plant Cell Culture Technology Section, Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, 400 085 Trombay, Mumbai, India.
Ganapathi
T R
TR
Srinivas
L
L
Revathi
C J
CJ
Bapat
V A
VA
D015153:Blotting, Western; D018929:Cell Culture Techniques; D002460:Cell Line; D003517:Cyclopentanes / Q000494:pharmacology; D018744:DNA, Plant / Q000032:analysis / Q000235:genetics; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000378:metabolism*; D054883:Oxylipins; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D020156:Salicylic Acid / Q000494:pharmacology; D014026:Nicotiana / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D014170:Transformation, Genetic
D003517:Cyclopentanes; D018744:DNA, Plant; D006514:Hepatitis B Surface Antigens; D054883:Oxylipins; D011993:Recombinant Fusion Proteins; C011006:jasmonic acid; D020156:Salicylic Acid