title: [Construction and sequencing of recombinant plasmid pcDNA3/GRA1 from Toxoplasma gondii].
abstract: OBJECTIVE
  To construct a mammalian expression plasmid pcDNA3/GRA1 to express dense granules antigen-1 (GRA1) of Toxoplasma gondii, and to lay a foundation for further studying the protective immunity of pcDNA3/GRA1 as a DNA vaccine.
  
  
  METHODS
  The GRA1 opening reading frame (ORF) was amplified with two specific primers. The ORF and plasmid pcDNA3 were digested with EcoR I and Xho I respectively and the ORF was ligated into the pcDNA3 at polylinker. The recombinant vector pcDNA3/GRA1 was characterized by PCR, restriction enzyme digestion, and sequencing analysis.
  
  
  RESULTS
  The expected ORF, 573 bp long, was amplified by PCR, and inserted into plasmid pcDNA3. PCR, restriction enzyme digestion and sequencing analysis showed that pcDNA3/GRA1 contained GRA1 ORF with the right orientation.
  
  
  CONCLUSIONS
  The mammalian expression vector pcDNA3/GRA1 is successfully constructed.
authors.affiliation: Department of Parasitology, Xiangya School of Medicine, Central South University, Changsha 410078, China.
authors.forename: Li-ting
authors.initials: LT
authors.lastname: Cai
authors.forename: Heng-ping
authors.initials: HP
authors.lastname: Shu
authors.forename: Li-ping
authors.initials: LP
authors.lastname: Jiang
mesh_terms: D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D017422:Sequence Analysis, DNA; D014122:Toxoplasma / Q000235:genetics*
chemical_list: D000953:Antigens, Protozoan; D004274:DNA, Recombinant; C412913:GRA1 antigen; D011993:Recombinant Fusion Proteins