中国脑型疟患者恶性疟原虫分离株裂殖子表面蛋白2基因的分子？…

目的 为设计研制安全有效的人脑型疟疫苗提供科学依据。方法 应用多聚酶链反应（ＰＣＲ）对５例中国脑型疟患者恶性朱虫云南省勐腊县勐罕（ＣＭＨ／ＹＮ）分离株和云南省盈江县农场（ＣＹＪ／ＹＮ）分离株基因组裂殖子表面蛋白２（ＭＳＰ２）基因进行扩增,将扩增产物分别经ＢａｍＨＩ和Ｈｉｎｄ Ⅲ双酶切后,回收的ＭＳＰ２基因分子定向克隆Ｍ１３ｍ１８和Ｍ１３ｍｐ１９载体,按Ｓａｎｇｅｒ双脱氧链终止法进行ＤＮＡ序列测定,

Molecular cloning and sequencing of genes encoding MSP2 isolates strains from two of Plasmodium falciparum from Chinese patients with cerebral malaria

OBJECTIVE: To provide the scientific evidence for designing safe and effective vaccines of human cerebral malaria. METHODS: Genomic DNA samples of two isolated Plasmodium falciparum isolate strains prepared directly from 5 cases of cerebral malaria patients' blood in mengla County, Yunnan Province (CMH/YN) and in Yingjiang County, Yunnan Province (CYJ/YN) were used for polymerase chain reaction (PCR) amplification and the two pairs of oligonucleotides for the highly conserved genes encoding FC27 merozoite surface protein 2 (MSP2) of Papua New Guinea strain of Plasmodium falciparum were used as primers. The PCR products were digested with BamH1 and Hind III respectively, and the generated fragment MSP2 were cloned into M13mp18 and M13mp19 vectors and their DNA was analyzed as the templates for DNA sequencing by the dideoxy chain-termination method. RESULTS: Compared with the published findings, FC27, K1, IC1 and CAMP sequences, DNA sequences of MSP2 from two isolated CMH/YN and CYJ/YN of Plasmodium falciparum strains from Chinese patients with cerebral malaria contained identical genes composed of 800 bp, encoding 264 amino acid, which were highly homologous up to 98.8% with that of FC27, K1 strain other than the IC1, CAMP strain. CONCLUSION: It is the first record of DNA sequencing of MSP2 determined from two isolated CMH/YN and CYJ/YN of Plasmodium falciparum strains from Chinese patients with cerebral malaria, MSP2 mutation may be one factor leading to the localized cerebral damage which causes clinical coma of human cerebral malaria.