title: [Introduction of Plasmodium falciparum C-terminal region of the merozoite surface protein gene into the chloroplast of tobacco].
abstract: OBJECTIVE
  To construct chloroplast expression vector, and introduce the C-terminal region of the merozoite surface protein 1 gene (msp1-42) of Plasmodium falciparum 3D7 strain into the chloroplast genome of tobacco for expression of the recombinant protein MSP1-42.
  
  
  METHODS
  Forward and reverse primers, adjusted to tobacco chloroplast codon preferences, were used for generation of msp1-42 gene from pBluntmsp plasmid which contains msp1-42 gene. A chloroplast expression vector LRrrmsp was constructed and bombarded into leaves of tobacco by a biolistic He particle delivery system. Media containing 500 mg/L spectinomycin were used for selection of spectinomycin resistant plant. PCR was carried out to check the introduction of the msp1-42 and aadA genes into the chloroplast genome. The transgenic plants with msp1-42 and aadA gene insertion were cut and cultured on the generation MS media containing 500 mg/L spectinomycin for at least 3 turns, and multiple PCR were applied to analyse their homogenization.
  
  
  RESULTS
  A chloroplast expression vector LRrrmsp was constructed and confirmed with PCR and enzyme digestion analysis. Six transformants were obtained with a transformation rate 0.6/gun. The msp1-42 and aadA genes were amplified from spectinomycin resistant plants by PCR detection. Wild type chloroplast gene was detected by multiple-PCR analysis.
  
  
  CONCLUSIONS
  A chloroplast expression vector containing msp1-42 gene was constructed. The msp1-42 gene was introduced into chloroplast genome of tobacco and heterogeneous transgenic tobacco was obtained.
authors.forename: Qin
authors.initials: Q
authors.lastname: Chen
authors.forename: Wan-Qi
authors.initials: WQ
authors.lastname: Liang
authors.forename: Bing-Jun
authors.initials: BJ
authors.lastname: Qian
authors.forename: Hui-Feng
authors.initials: HF
authors.lastname: Shen
authors.forename: Jian-Ping
authors.initials: JP
authors.lastname: Cao
authors.forename: Yu-Xin
authors.initials: YX
authors.lastname: Xu
authors.forename: Da-Bing
authors.initials: DB
authors.lastname: Zhang
authors.forename: Lin-Hua
authors.initials: LH
authors.lastname: Tang
mesh_terms: D002736:Chloroplasts / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors; D020066:Merozoite Surface Protein 1 / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics; D010957:Plasmids; D010963:Plasmodium falciparum / Q000235:genetics*; D014026:Nicotiana / Q000235:genetics*
chemical_list: D020066:Merozoite Surface Protein 1