棘阿米巴分离株的分子生物学特性研究

Study of molecular biological characterization of Acanthamieba spp.isolates

[目的]研究土壤和淡水中分离的3种棘阿米巴分离株分子生物学特性.[方法]提取棘阿米巴株CJY/S3,CG/S1和CJY/W1基因组18SrDNA,行PCR扩增后测定序列,与T1~T13型序列进行比较分析;提取线粒体DNA用EcoRⅠ酶切,观察限制性片段长度多态性（RFLP）.[结果]3种棘阿米巴分离株CJY/S3,CG/S1和CJY/W1的18S rDNA大小分别为2 292,2 292,2 252bp.CG/S1和CJY/S3的基因型为T5,基因序列接近于A.lenticulata 68-2;CJY/W1的基因型为T13,基因序列接近于Acanthamoeba sp.U/HC1株.CJY/S3和CG/S1株显示相同的线粒体DNA片段模式,不同于CJY/W1株.[结论]棘阿米巴CJY/W1株与CJY/S3,CG/S1株具有不同的分子生物学特性.

OBJECTIVE To study the molecular biological characterization of three strains of Acanthamieba spp.isolated from tap water and soil.METHODS 18 SrDNA genome of Acanthamieba spp.CG/S1,CJY/S3,CJY/W1 strains were extracted,and the sequences were detected after PCR amplification and were compared with sequences of genotypes T1-T13.Extracted mitochondrial DNA was digested with EcoRⅠ and the Mt-DNA-RFLP were observed.RESULTS The full length of 18 SrDNA genome of CG/S1,CJY/S3 and CJY/W1 were 2 292,2 292 and 2 252 bp,respectively.The 18 SrDNA genotype of CG/S1 and CJY/S3 was T5,and the gene sequences were close to Acanthamieba lenticulata 68-2.The 18 SrDNA genotype of CJY/W1 was T13,and the gene sequences was close to Acanthamieba spp.U/HC1.Pattern of Mt-DNA-RFLP of CJY/W1 was differences with strains of CG/S1 and CJY/S3.CONCLUSION The Acanthaoeba CJY/W1 strain isolated from tap water has different molecular biological characterization with CB/S1 and CJY/S3 strains isolated from soil.