In vitro amplification of Toxoplasma gondii rhoptry protein 2 gene and construction of its eukaryotic expression plasmid.
OBJECTIVE
To construct a recombinant eukaryotic expression plasmid containing rhoptry protein 2 (ROP2) gene of Toxoplasma gondii.


METHODS
The truncated ROP2 gene was amplified from the genomic DNA of Toxoplasma gondii RH strain and cloned into plasmid pGEX-4T-1. The PCR product was subcloned into an eukaryotic expression plasmid pcDNA3 which was identified by enzyme digestion and PCR amplification. Sequence analysis of the insert in the recombinant plasmid was performed by Sanger's method.


RESULTS
The amplified DNA fragment was about 1043 bp in length, with identical sequence with the published ROP2 gene sequence. Enzyme digestion and PCR analysis showed that the truncated ROP2 gene had been successfully cloned into plasmid pGEX-4T-1 and pcDNA3.


CONCLUSIONS
The truncated ROP2 gene of Toxoplasma gondii has been amplified and cloned into the eukaryotic expression plasmid pcDNA3.
Wu
Kun
K
Department of Parasitology, First Military Medical University, Guangzhou 510515, China. wukun@fimmu.com
Chen
Xiao-Guang
XG
Li
Hua
H
Zhi
Guo-Zhou
GZ
D000818:Animals; D001483:Base Sequence; D016054:DNA, Protozoan / Q000032:analysis; D005057:Eukaryotic Cells / Q000378:metabolism; D005784:Gene Amplification; D015870:Gene Expression; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000235:genetics*; D014122:Toxoplasma / Q000235:genetics*; D014162:Transfection
D016054:DNA, Protozoan; D008565:Membrane Proteins; D015800:Protozoan Proteins; C090555:ROP 2 protein, Toxoplasma gondii