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首页> 外文期刊>Infection and immunity >Codon Optimization of Gene Fragments EncodingPlasmodium falciparum Merzoite Proteins Enhances DNA Vaccine Protein Expression and Immunogenicity in Mice
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Codon Optimization of Gene Fragments EncodingPlasmodium falciparum Merzoite Proteins Enhances DNA Vaccine Protein Expression and Immunogenicity in Mice

机译：编码恶性疟原虫裂殖子蛋白质的基因片段的密码子优化增强了小鼠中DNA疫苗蛋白的表达和免疫原性

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In contrast to conventional vaccines, DNA and other subunit vaccines exclusively utilize host cell molecules for transcription and translation of proteins. The adenine plus thymine content of Plasmodium falciparum gene sequences (～80%) is much greater than that of Homo sapiens(～59%); consequently, codon usage is markedly different. We hypothesized that modifying codon usage of P. falciparumgenes encoded by DNA vaccines from that used by the parasite to those resembling mammalian codon usage would lead to increased P. falciparum protein expression in vitro in mouse cells and increased antibody responses in DNA-vaccinated mice. We synthesized gene fragments encoding the receptor-binding domain of the 175-kDaP. falciparum erythrocyte-binding protein (EBA-175 region II) and the 42-kDa C-terminal processed fragment of the P. falciparum merozoite surface protein 1 (MSP-142) using the most frequently occurring codon in mammals to code for each amino acid, and inserted the synthetic genes in DNA vaccine plasmids. In in vitro transient-expression assays, plasmids containing codon-optimized synthetic gene fragments (pS plasmids) showed greater than fourfold increased protein expression in mouse cells compared to those containing native gene fragments (pN plasmids). In mice immunized with 0.5, 5.0, or 50 μg of the DNA plasmids, the dose of DNA required to induce equivalent antibody titers was 10- to 100-fold lower for pS than for pN plasmids. These data demonstrate that optimizing codon usage in DNA vaccines can improve protein expression and consequently the immunogenicity of gene fragments in DNA vaccines for organisms whose codon usage differs substantially from that of mammals.
机译：与常规疫苗相反，DNA和其他亚基疫苗仅利用宿主细胞分子进行蛋白质的转录和翻译。恶性疟原虫基因序列中腺嘌呤和胸腺嘧啶的含量（〜80％）远大于智人的同源性（〜59％）。因此，密码子用法明显不同。我们假设修改 P的密码子用法。由DNA疫苗编码的恶性疟原虫基因，与寄生虫所使用的类似，类似于哺乳动物密码子的用法，会导致 P增加。小鼠细胞中falciparum 蛋白的体外表达和接种DNA的小鼠中抗体应答的增加。我们合成了编码175-kDaemP的受体结合结构域的基因片段。恶性疟原虫红细胞结合蛋白（EBA-175区域II）和 P的42 kDa C末端加工片段。恶性疟原虫的裂殖子表面蛋白1（MSP-1 42 ）利用哺乳动物中最常见的密码子编码每个氨基酸，并将合成基因插入DNA疫苗质粒中。在体外瞬时表达测定中，与含有天然基因片段（pN质粒）的质粒相比，含有密码子优化的合成基因片段（pS质粒）的质粒在小鼠细胞中的蛋白质表达增加了四倍以上。在用0.5、5.0或50μgDNA质粒免疫的小鼠中，与pN质粒相比，pS诱导等效抗体滴度所需的DNA剂量要低10到100倍。这些数据表明，优化DNA疫苗中密码子的使用可以改善蛋白质表达，从而改善DNA疫苗中基因片段对那些密码子使用与哺乳动物有很大不同的生物的免疫原性。

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