[Construction of the plant expression vectors containing the multiepitope gene of Toxoplasma gondii].
OBJECTIVE
To construct the plant expression vectors containing the multiepitope gene of Toxoplasma gondii (TGMG).


METHODS
1. TGMG was subcloned into pBAC55 vector to construct the intermediate plasmid pB35MG. The E35S/TGMG/NOS3' fragment was cleaved from pB35MG and ligated into the plant binary vector pCAMBIA2300 to construct the plant expression vector pC35MG. 2. Tomato fruit-specific E81. 1 promoter was introduced to pB35MG to construct pB35E1MG vector. The E35SE81. 1/TGMG/NOS3' fragment was subcloned into pCAMBIA2300 to construct the plant expression vector pC35E1MG. 3. Tomato fruit-specific E82.2 promoter was inserted to pB35MG to construct pBE2MG vector. The E82.2/TGMG/NOS3' fragment was subcloned into pCAMBIA2300 to construct the plant expression vector pCE2MG. The insert gene TGMG in the vectors pB35MG, pC35E1MG and pCE2MG were confirmed by sequencing. 4. pC35MG, pC35E1MG and pCE2MG were introduced into Agrobacterium tumefaciens strain LBA4404 competent cell.


RESULTS
Digestion with restriction enzymes proved that all recombinant vectors had the inserts with expected length of the target fragments. And the sequencing results were confirmed correct.


CONCLUSIONS
The TGMG intermediate vectors pB35MG, pB35E1MG and pBE2MG and the plant expression vectors pC35MG, pC35E1MG and pCE2MG were constructed successfully, and the three plant expression vectors were introduced into Agrobacterium tumefaciens.
Zhou
Xiao-hong
XH
Department of Parasitology, First Military Medical University, Guangzhou 510515.
Chen
Xiao-guang
XG
Zhang
Xiao-dong
XD
Yang
Pei-liang
PL
Xi
Jia-fei
JF
Hu
Jian-jun
JJ
Wang
Ya-nan
YN
Li
Lin
L
Shen
Shu-man
SM
D016960:Agrobacterium tumefaciens / Q000235:genetics; D000818:Animals; D003001:Cloning, Molecular; D000939:Epitopes / Q000235:genetics; D015870:Gene Expression; D017125:Genes, Protozoan; D005822:Genetic Vectors; D018551:Solanum lycopersicum / Q000235:genetics*; D030821:Plants, Genetically Modified; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014122:Toxoplasma / Q000235:genetics*
D000939:Epitopes