关键词: 恶性疟原虫, 叶绿体, 基因表达, 裂殖子表面蛋白1, 植物疫苗

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 transformmants 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. Conclusion A chloroplast expression vector containing msp1-42 gene was constructed. The msp1-42 gene was intro-duced into chloroplast genome of tobacco and heterogenous transgenic tobacco was obtained.

Key words: Plasmodium falciparum, Chloroplast, Gene expression, Merozoite surface protein 1, Plant vaccine