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。
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.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
1993年第2期81-85,共5页
Chinese Journal of Parasitology and Parasitic Diseases
基金
国家自然科学基金 No.3907053
关键词
间日疟原虫
聚合酶链反应
分子克隆
Plasmodium vivax
small subunit ribosomal RNA gene
polymerase chain reaction
molecular cloning
DNA sequence analysis
