[Construction and expression of the eukaryotic coexpression plasmid containing Ag85B gene of Mycobacterium tuberculosis and IL-12 gene].
OBJECTIVE
To construct an eukaryotic coexpression plasmid containing Mycobacterium tuberculosis (MTB) Ag85B and IL-12 genes.


METHODS
MTB Ag85B gene and IL-12 gene were cloned into pBudCE4.1 which has multiple promoters to construct recombinant plasmid pBud85B-IL12. The recombinant plasmid was transfected into COS-7 cells and the expression of target genes was assessed by RT-PCR and ELISA.


RESULTS
The expressions of Ag85B and IL-12 could be detected in COS-7 cells.


CONCLUSIONS
The recombinant plasmid pBud85B-IL12 was constructed and expressed successfully, which lays the foundation for further development of DNA vaccine against tuberculosis.
Jiang
Shan
S
Department of Microbiology, Chongqing University of Medical Sciences, Chongqing 400016, China. meimei0075@sina.com.cn
Zhu
Dao-yin
DY
Luo
Xu-dong
XD
Chen
Quan
Q
Jiang
Ying
Y
D000217:Acyltransferases / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D019556:COS Cells; D002522:Chlorocebus aethiops; D018664:Interleukin-12 / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D011994:Recombinant Proteins; D018664:Interleukin-12; D000217:Acyltransferases; C121433:antigen 85B, Mycobacterium tuberculosis