中国寄生虫学与寄生虫病杂志 ›› 1995, Vol. 13 ›› Issue (3): 203-208.

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恶性疟原虫云南勐捧分离株裂殖子顶端膜抗原Ⅰ序列分析

张龙兴,詹斌,王聚君,冯晓平   

  1. 中国预防医学科学院寄生虫病研究所,上海; 200025; 第二军医大学寄生虫学教研室; 上海; 200433; 中国预防医学科学院寄生虫病研究所,上海; 200025; 中国预防医学科学院寄生虫病研究所,上海; 200025; 中国预防医学科学院寄生虫病研究所,上海; 200025
  • 出版日期:1995-08-31 发布日期:2017-01-05

SEQUENCE ANALYSIS OF APICAL MEMBRANE ANTIGEN Ⅰ FROM A PLASMODIUM FALCIPARUM ISOLATE COLLECTED FROM MENGPENG TOWNSHIP, YUNNAN PROVINCE

  • Online:1995-08-31 Published:2017-01-05

摘要: 根据文献报道的恶性疟原虫裂殖子顶端膜抗原IDNA序列的保守区,设计并合成两对20聚寡核苷酸作引物,对恶性疟原虫勐捧分离株DNA进行PCR扩增。扩增产物经BamHI和EcoRI双酶解后,克隆入具有相应末端的M13mp8和M13mp8载体,转化JPA101,生长于含X-gal的培基上,抽提无色噬菌斑DNA,以DNA序列测定仪进行DNA序列测定,并自动翻释成氨基酸序列。用双脱氧终止法测定部分DNA序列,并与DNA序列测定仪测定的序列进行比较。结果表明,扩增的序列长度为1773个碱基,编码591个氨基酸。与参照序列比较,有17个点突变,引起15个密码子的取代,其中,除1个密码子为同义取代外,其余密码子均为非同义取代,造成14个氨基酸的取代。点突变在序列中呈散在分布,但在氨基酸序列中第160-210位相对较集中。

关键词: 恶性疟原虫, 裂殖子, 顶端膜抗原Ⅰ, DNA序列分析

Abstract: Genomic DNA from a Plasmodium falciparum isolate (CMP-1) collected from a falciparum malaria patient at Mengpeng Township, Mengla County, Yunnan Province was used as the template for PCR amplification primered with oligonucleotides from the conserved regions of the apical membrane antigen I (AMA-1) as reported. PCR products recovered from the low melting-point agarose gel electrophoresis were digested with BamHI and EcoRI to generate fragments approximately 420, 445 and 900 bp which were used as the inserts to insert into the M13 mp8 and M13 mp9 vectors for sequencing using 373 sequencer. 1773 bases were read out, among which 17 point mutations were found to result in substitutions of 15 codens which were non-synonymous mutations except for one as compared with that of the consensus. In particular, a relatively hot region of variation was apparent between amino acids 160 and 210.

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