恶性疟原虫核糖体小亚基rRNA部分基因片段的克隆及序列分析

Cloning and analysis partial squence of SSUrRNA gene of Plasmodium falciparum

目的了解我国恶性疟原虫云南地理株核糖体小亚基rDNA的核苷酸序列。方法根据已知恶性疟原虫核糖体小亚基rDNA序列．在其基因的中下游分别设计引物，采用聚合酶链式反应（PCR）技术，从云南省来源的恶性疟体外培养原虫的基因组DNA中扩增出的基因片段酶切后将其插入pUC118／119载体，用Biosystems373ADNA序列分析仪测定。结果我国恶性疟原虫云南地理株核糖体小亚基rDNA核苷酸序列，与巴西IMTM22／7G8株的序列比较，在该基困的第1563位有一个A被T替换。结论该突变点位于真核生物核糖作小亚基rRNA基因已知序列种类的第三个可变区域内。

Sequencing of SSUrRNA gene fragment at 1450-1622 from genomic DNA Plasmodium falciparum from Hainan.Method One 172 bp SSUrRNA gene fragment at 1450-1622 from genomic DNAPlasmodium falciparum from Hainan province P.R. China, was amplified with polymerase chain reaction (PCR). After digestion the amplified DNA fragment of Plasmodium falciparum were inserted into vector pUC118/119, then the seqence was determined by dideoxy- nucleotide terminator method and analysed with computer programs.Results The results show that the sequence is the same as that of P. falciparum SSUrDNA in 7G8 strain from IMTM22 substitutiory except for one T instead of A at position 1563.Conclusions This mutated region locates in the third variable region of eukaryote SSUrRNAgene.