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目的 了解恶性疟原虫海南株 (FCC1/HN)乳酸脱氢酶基因全编码区核苷酸序列变异情况。方法 PCR扩增恶性疟原虫海南株 (FCC1/HN)LDH基因全编码区 ,产物经EcoRⅠ +SalⅠ酶切后 ,定向克隆至pGEX - 4T - 1表达质粒 ,采用Sanger双脱氧链末端终止法测序 ,与其它生物LDH进行同源性分析。 结果 成功地扩增了恶性疟原虫海南株 (FCC1/HN)LDH编码基因 ,大小为 95 1bp ,无内含子 ,与Honduras株LDH基因相比 ,两者有 5个碱基不同 ,推导的氨基酸序列有 4个氨基酸残基不同 ,与刚地弓形虫、隐孢子虫和芽孢杆菌的同源性分别为 49.0 6 % (15 6 / 318)、42 .5 8% (132 / 310 )、31.6 1% (98/310 )。与人的LDH -A、LDH -B和LDH -C的同源性分别为 30 .19% (93/ 30 8)、2 9.5 5 % (91/ 30 8)和 33.44 % (10 4/ 311)。结论 FCC1/HN株与Honduras株LDH高度同源 ,疟原虫LDH明显不同于其它生物LDH
Objective To understand the variation of the nucleotide sequence of the complete coding region of lactate dehydrogenase gene in Plasmodium falciparum Hainan (FCC1 / HN). Methods The complete coding region of LDH gene of Plasmodium falciparum (FCC1 / HN) was amplified by PCR. The product was digested with EcoR Ⅰ + Sal Ⅰ and cloned into pGEX - 4T - 1 expression plasmid. The sequencing was performed by Sanger dideoxy chain termination method. Homology analysis with other organisms LDH. Results The LDH encoding gene of P. falciparum Hainan strain (FCC1 / HN) was successfully amplified with a size of 951 bp and no introns. Compared with the LDH gene of Honduras strain, the two genes had 5 bases, and the deduced amino acids The sequences had 4 amino acid residues, which were 49.0 6% (15 6/318), 42.58% (132/310) and 31.6 1 % (98/310). The homologies with human LDH-A, LDH-B and LDH-C were 30.19% (93/308), 29.5% (91/308) and 33.44% (104/1191), respectively . Conclusion The FCC1 / HN strain is highly homologous to the Honduras strain LDH, and the Plasmodium LDH is significantly different from other biological LDH