[Expression of glycoprotein gene of the rabies virus 3aG strain by E-3 deleted adenovirus recombinant].
OBJECTIVE
To analyze Chinese rabies virus vaccine strain 3aG glycoprotein (GP) gene and further produce GP by E3-deleted human adenovirus recombinant.


METHODS
Chinese rabies virus vaccine strain 3aG glycoprotein gene was cloned by RT-PCR and its sequence was determined by DNA sequencing. Cotransfection was performed to obtain adenovirus recombinant. The expressed glycoprotein was examined by ELISA and its immunogenicity was evaluated by testing neutralizing antibody level of mice inoculated with the recombinant virus.


RESULTS
DNA sequencing showed that the open reading frame of GP gene contains 1,575 nucleotides and five of the deduced amino acids are different from the previous report. The recombinant adenovirus containing GP gene in E3 region was obtained by cotransfecting 293 cells and rounds of plaque purification. ELISA assay demonstrated that the GP gene can be efficiently expressed and rapid fluorescent focus inhibition test (RFFTT) showed it can also elicit GP specific neutralizing antibody.


CONCLUSIONS
Chinese rabies virus vaccine strain 3aG glycoprotein was successfully expressed by E3-deleted human adenovirus recombinant and specific neutralizing antibody can be elicited in mice after immunization by the recombinant.
Zhang
X
X
Institute of Basic Medical Science, Chinese Acdemy of Medical Scienee & Peking Union Medical College, Beijjng 100005, China.
Zhang
Y
Y
Li
W
W
D017503:Adenovirus E3 Proteins / Q000235:genetics; D000260:Adenoviruses, Human / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D000917:Antibody Formation; D000956:Antigens, Viral; D002478:Cells, Cultured; D005260:Female; D015870:Gene Expression; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics*; D051379:Mice; D008815:Mice, Inbred Strains; D011819:Rabies Vaccines / Q000276:immunology; D011820:Rabies virus / Q000235:genetics* / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics*
D017503:Adenovirus E3 Proteins; D000956:Antigens, Viral; D006023:Glycoproteins; D011819:Rabies Vaccines; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; C079096:adenovirus 5, rabies glycoprotein recombinant vaccine; C064951:glycoprotein G, Rabies virus