中国寄生虫学与寄生虫病杂志 ›› 1993, Vol. 11 ›› Issue (2): 81-85.

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云南省间日疟原虫SSUrRNA基因片段的体外扩增、克隆及序列分析

万磊,智刚,陈培霞,薛采芳,姜绍谆,刘珺   

  1. 第四军医大学寄生虫学教研室; 第四军医大学药物研究所; 第四军医大学寄生虫学教研室; 第四军医大学寄生虫学教研室; 第四军医大学微生物学教研室; 第四军医大学寄生虫学教研室
  • 出版日期:1993-05-31 发布日期:2017-01-06
  • 基金资助:
    国家自然科学基金 No.3907053

AMPLIFICATION, CLONING AND SEQUENCE ANALYSIS OF A SSUrRNA GENE FRAGMENT OF PLASMODIUM VIVAX ISOLATES FROM YUNNAN PROVINCE

  • Online:1993-05-31 Published:2017-01-06

摘要: 根据已知间日疟原虫、其它相关原虫及人小亚单位核糖体核糖核酸(SSUrRNA)基因序列,借助计算机程序分析,设计一对寡聚核苷酸引物。采用聚合酶链式反应(PCR)技术,从云南省两例间日疟患者血样DNA抽提物中,均扩增出长约641个碱基对(bp)的SSUrRNA基因特定片段;双脱氧链末端终止法测序结果表明,两份样本扩增片段DNA序列完全相同,但与背景序列比较,在第269位出现碱基置换由G变为A,而在第630位则缺失一碱基C,从而导致该两处限制性内切酶位点的改变。

关键词: 间日疟原虫, 核糖体核糖核酸基因, 聚合酶链式反应, 分子克隆, DNA序列分析

Abstract: According to known SSUrUNA sequences ot Plasmodium vivax , correlated protozoa and human being, sequences of oligonucleotide primers were defined with computer programming. Specific SSUrDNA fragment of P. vivax , about 640 bp in length, was directly amplified by two temperature point polymerase chain reaction from extracted genomic DNA of two blood samples of vivax malaria patients from Yunnan Province. Using dideoxynu-cleotide terminator method, the sequences of amplified DNA fragments were determined separately and showed no difference between the two samples. However, comparison of the sequence reported by Waters AP and McCutchan TF(1989)and that of amplified fragment of Yunnan P. vivax isolates revealed the existence of nucleotide substitution and deletion which occured respectively in the sites 269 and 630,and resulted in the change of restriction map.