[Cloning and identification of ROP2 gene of Toxoplasma gondii].
OBJECTIVE
To amplify ROP2 from the genomic DNA of Toxoplasma gondii RH strain and construct eukaryotic expression plasmid pc-DNA3-ROP2.


METHODS
Tachyzoites of T. gondii RH strain were collected and depurated to obtain genome. A pair of primers was designed and synthesized according ROP2 gene sequence. The gene fragment encoding ROP2 was amplified from the genomic DNA of T. gondii RH strain by means of PCR. It was then reclaimed and purified, and inserted into cloning vector pUCm-T. The recon was cut by EcoR I, Hind III, and the inserted ROP2 gene fragment was subcloned into pc-DNA3 eukaryotic expression vector using T4DNA ligase, followed by further PCR identification, double digestion via restrictive enzymes, and sequencing.


RESULTS
The amplified specific gene fragment of ROP2 was about 1.7 kb in length. The gene fragment cloned and subcloned into pc-DNA3 was correct, and the eukaryotic expression plasmid contained ROP2 gene fragment was successfully constructed.


CONCLUSIONS
The recombinant plasmid pc-DNA3-ROP2 was successfully constructed.
Wei
Qing-kuan
QK
Shandong Institute of Parasitic Diseases, Jining 272033, China.
Zhang
Dian-bo
DB
Li
Jin
J
Li
Gui-ping
GP
Wang
Hong-fa
HF
Cui
Yong
Y
Bai
Xue-lian
XL
Fu
Bin
B
Liu
Yu-bing
YB
Zai
De-fu
DF
Huang
Bing-cheng
BC
Liu
Ke-yi
KY
Han
Guang-dong
GD
D000818:Animals; D016054:DNA, Protozoan; D015870:Gene Expression; D008565:Membrane Proteins / Q000235:genetics* / Q000302:isolation & purification; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000235:genetics* / Q000302:isolation & purification; D011994:Recombinant Proteins; D014122:Toxoplasma / Q000235:genetics* / Q000276:immunology
D016054:DNA, Protozoan; D008565:Membrane Proteins; D015800:Protozoan Proteins; C090555:ROP 2 protein, Toxoplasma gondii; D011994:Recombinant Proteins