title: Construction and identification of Complex DNA vaccine of hepatitis B and Toxoplasma gondii.
abstract: OBJECTIVE
  To construct and identify multi-gene recombinant expression vector pcDNA3-HBsAg-p30-ROP2.
  
  
  METHODS
  Primers were designed according to the gene sequences of restriction enzyme cutting site of recombinant pcDNA3-p30-ROP2 and hepatitis B surface antigen (HBsAg). The target fragment of HBsAg was amplified and cloned to expression vector pcDNA3-p30-ROP2 by restriction enzyme digestion and ligation. The recombinant expression vector pcDNA3-HBsAg-p30-ROP2 was identified by PCR detection, followed by enzyme restriction and sequencing.
  
  
  RESULTS
  The target fragment of HBsAg was successfully amplified, and the multi-gene eukaryotic expression vector pcDNA3-HBsAg-p30-ROP2 was established. PCR detection and restriction enzyme digestion showed that the length of the target fragment was consistent with the theoretical value. The recombinant expression vector contained the complete sequences of p30-ROP2 compound gene and HBsAg.
  
  
  CONCLUSIONS
  Multi-gene recombinant expression vector pcDNA3-HBsAg-p30-ROP2 was successfully established. The constructed expression vector could be used to develop multi-gene nucleic acid vaccines.
authors: Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Disease Jining 272033, China.
authors: Qing-Kuan
authors: QK
authors: Wei
authors: Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Disease Jining 272033, China.
authors: Ting
authors: T
authors: Xiao
authors: Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Disease Jining 272033, China.
authors: Jin
authors: J
authors: Li
authors: Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Disease Jining 272033, China.
authors: Kun
authors: K
authors: Yin
authors: Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Disease Jining 272033, China.
authors: Feng-Ju
authors: FJ
authors: Jia
authors: Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Disease Jining 272033, China.
authors: Chao
authors: C
authors: Xu
authors: Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Disease Jining 272033, China.
authors: Gui-Hua
authors: GH
authors: Zhao
authors: Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Disease Jining 272033, China.
authors: Yong
authors: Y
authors: Cui
authors: Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Disease Jining 272033, China.
authors: Gong-Zhen
authors: GZ
authors: Liu
authors: Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Disease Jining 272033, China.
authors: Hui
authors: H
authors: Sun
authors: Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Disease Jining 272033, China.
authors: Hong-Tao
authors: HT
authors: Jiang
authors: Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Disease Jining 272033, China.
authors: Ge
authors: G
authors: Yan
authors: Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Disease Jining 272033, China.
authors: Bing-Cheng
authors: BC
authors: Huang
keywords: HBsAg; Rhoptry protein 2 (ROP2); Toxoplasma gondii; genetic recombination; surface antigen 1 (p30)