化学合成恶性疟原虫杂合45肽抗原基因在大肠杆菌中的表达

A PEPTIDE OF 45 AMINO-ACID RESIDUES ENCODED BY A SYNTHETIC HYBRID GENE OF PLASMODIUM FALCIPARUM EXPRESSED IN ESCHERICHIA COLI

为了使化学合成的人恶性疟原虫杂合45肽抗原基因(HPFAG)在大肠杆菌中表达,我们将HPFAG与表达载体pWR450.1质粒重组连接,转化大肠杆菌,获得一批转化子。这些转化子经质粒DNA斑点杂交试验,蛋白质SDS—PAGE和DNA酶解分析,表明化学合成的HPFAG与表达载体质粒发生了重组,且有6个重组子在大肠杆菌中与p—半乳糖苷酶呈融合蛋白形式表达。

Based on the paper of Manual E. Patarroyo(1988) ,'a synthetic vaccine protects human against challenge with asexual blood stages of Plasmodium faldparum ',We deduced the nucleotide-sequencing and synthesized a hybrid antigen gene encoding a peptide of 45 amino-acid residues. In order to be expressed,the gene was recombined with a plasmid vector ,pWR450. 1, and transformed into E. coli . The recombinant clones were screened with Dot blot hybridization and protein SDS-PAGE. The recombinant plasmid DNA was digested by restriction endonucle-ases for analysis. The results showed that the gene recombined with the plasmid and expressed as a fusion protein with β-galactosidase in E. coli ,Its immunogentcity has been confirmed by animal serological experiment.