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试验证实我国黑热病不同疫区分离出的 5个L d 分离株的小亚基核糖体DNA可变区序列存在点突变。方法 将 7个利什曼原虫种 /株的nDNA进行PCR扩增 ,将扩增出的SSUrDNA基因的特异片段克隆于pGEMR TEasyVector上 ,采用通用引物M1 3进行PCR扩增 ,全自动测序仪测序。结果 序列分析显示 ,扩增的 7种 /株利什曼原虫SSUrDNA序列大小为 3 92bp ,5个点突变均发生在两个独特序列区 (UQ I和UQ II) ;无移码突变。经与基因库中利什曼原虫序列类似性比较 ,同源性在 98%以上。结论 首次报道我国黑热病不同疫区的 5个L d 分离株的SSUrDNA可变区存在 5个点突变。
The experiment confirmed that there were point mutations in the small subunit ribosomal DNA variable region sequences of 5 L d isolates isolated from different epidemic areas in China. Methods The nDNA of seven Leishmania species / strains were amplified by PCR. The specific fragment of SSU rDNA gene was cloned into pGEMR TEasyVector. PCR was carried out by universal primer M1 3 and sequenced by automatic sequencing. Results The sequence analysis showed that the size of SSUrDNA of 7 species / strains of Leishmania amplified was 3 92bp, and all five point mutations occurred in two unique sequence regions (UQ I and UQ II); there was no frameshift mutation. The similarity of Leishmania sequences in the gene pool was over 98%. Conclusions For the first time, there are 5 point mutations in the SSUrDNA variable region of 5 L d isolates from different epidemic areas in China.