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目的 探索埃及血吸虫基因组内是否存在曼氏血吸虫基因组“特异的”高度重复序列Sm1- 7。方法 以埃及血吸虫基因组 DNA为模板 ,用根据 Sm1- 7设计并合成的寡核苷酸引物进行多聚酶链式反应 (PCR)扩增 ,选取扩增产物中长约 30 0 bp的 DNA片段 ,经过预处理后与去磷酸化的质粒载体 p Bluescript (KS+)进行连接 ,连接产物转化至大肠杆菌 JM10 9。重组质粒经纯化后作DNA序列测定。结果 从埃及血吸虫基因组 DNA中成功地扩增出与曼氏血吸虫基因组重复序列Sm1- 7同源的重复序列。通过 DNA序列测定显示 ,除个别碱基不同外 ,从埃及血吸虫基因组获得的重复序列与曼氏血吸虫基因组重复序列 Sm1- 7完全相同。灵敏度分析显示该重复序列在埃及血吸虫基因组内的拷贝数远低于其在曼氏血吸虫基因组内的拷贝数。结论 曼氏血吸虫基因组“特异的”DNA重复序列 Sm1- 7在埃及血吸虫基因组内同样存在
Objective To explore the presence of a “specific” highly repetitive sequence Sm1-7 in the genome of Schistosoma mansoni genome. Methods The genomic DNA of Schistosoma japonicum was used as a template to amplify the DNA fragment of 30 bp in the amplified product by polymerase chain reaction (PCR) with oligonucleotide primers designed and synthesized according to Sm1-7. After treatment, the dephosphorylated plasmid vector p Bluescript (KS +) was ligated and the ligation product was transformed into E. coli JM109. Recombinant plasmids were purified for DNA sequencing. As a result, a repeat sequence homologous to the Sm1-7 genome of Schistosoma mansoni was successfully amplified from the genomic DNA of S. japonicum. DNA sequence analysis revealed that the sequence repeats from the S. was identical to the S. aureus genome sequence Sml-7, except for individual bases. Sensitivity analysis showed that the copy number of this repeat within the genome of Schistosoma mansoni was much lower than its copy number in the S. mansoni genome. Conclusions The “specific” DNA repeat Sm1-7 of the Schistosoma mansoni genome is also present in the genome of Schistosoma mansoni