Abstract:

Objective   To construct, clone and identify the DNA recombinant plasmid pBudCE4.1-AMA1-ROP18. Methods   Amplify the target genes by PCR, construct the eukaryotic recombinant plasmid pBudCE4.1-AMA1-ROP18 and express it in human foreskin fibroblast (HFF) cells by transfection. Then identify the plasmid with SDS-PAGE and betaactin. Results   We proved that the recombinant plasmid pBudCE4.1-AMA1-ROP18 was constructed correctly by identification of sequencing, double enzyme digestion and PCR.We proved that AMA1 gene and ROP18 gene both transcribed and expressed correctly in mammalian cells by identification of RT-PCR and SDS-PAGE. Conclusion   The recombinant plasmid pBudCE4.1-AMA1-ROP18 is constructed successfully and expressed in HFF cells stably.

Key words: Toxocara; Plasmids；Cell transfection；Apical membrane antigen 1; Rhoptry protein 18