Concomitant cellular expression of heat shock regulated genes of hepatitis B virus surface antigen and of human growth hormone by a NIH-3T3 cell line.
A plasmid carrying a DNA fragment of hepatitis B virus, coding for the pre-S2 and the entire S region of the surface antigen (HBsAg), placed under the control of the promoter of the human 70 kDa heat shock protein gene (hsp70), was introduced into Line 6, a recombinant cell line that was selected from NIH-3T3 cells previously transfected with a similar construct coding for the human growth hormone cDNA gene (chGH) and with the plasmid pEJ carrying the Ha-rasEJ activated cellular oncogene. The resulting cell line, EMS8, expressed: (1) hsp70/HBsAg and hsp70/hGH hybrid genes, (2) the human Ha-rasEJ oncogene, and (3) the neomycin resistance gene, the two last plasmid markers being used for cell selection. EMS8 cells were able to carry out post-translational modifications of the middle M and the major S envelope proteins of HBV, such as assembly and glycosylation. Accordingly, the cells synthesized and secreted both free and glycosylated M and S viral proteins, and the human growth hormone protein. In addition concomitant expression of HBsAg and hGH proteins as well as their mRNA were detected in EMS8 cells at least up to 72 hr after heat induction instead of 24 hr in the case of hGH in Line 6 cells.
Department of Genetic Engineering, IntraCel S.A., Geneva-Carouge, Switzerland.
P
P
L'Hote
S
S
Alouani
J B
JB
Marq
F
F
Montandon
M
M
Chessebeuf-Padieu
M
M
Dreano
D016475:3T3 Cells; D000818:Animals; D002460:Cell Line; D013006:Growth Hormone / Q000096:biosynthesis / Q000235:genetics*; D006360:Heat-Shock Proteins / Q000235:genetics*; D006514:Hepatitis B Surface Antigens / Q000235:genetics*; D006801:Humans; D006824:Hybridization, Genetic; D051379:Mice; D010442:Peptide Chain Initiation, Translational / Q000502:physiology; D012333:RNA, Messenger / Q000032:analysis; D011994:Recombinant Proteins / Q000096:biosynthesis
D006360:Heat-Shock Proteins; D006514:Hepatitis B Surface Antigens; D012333:RNA, Messenger; D011994:Recombinant Proteins; D013006:Growth Hormone