弓形虫非同源性不同地理株的GRA1基因体外扩增片段的鉴定及比较研究

COMPARISON OF GRA1 GENE AMPLIFIED IN VITRO FROM DIFFERENT GEOGRAPHICAL AND HETEROLOGOUS TOXOPLASMA GONDII ISOLATES

目的对弓形虫人源性RH株、人源性ZS_2株、猪源性CN株3个不同地理株的GRA1基因片段鉴定并比较异同。方法采用PCR技术和限制性内切酶技术，分别对弓形虫3株的GRA1基因片段进行体外扩增。扩增的目的基因片段分别用3种内切酶HindⅢ、HPaⅡ、TaqⅠ进行单酶切鉴定、比较。结果从弓形虫3个分离株RH株、ZS_2株、CN标基因组DNA中扩增出785bp的GRA1基因片段。3个株的GRA1基因分别用3种内切酶酶切后，酶切片段与理论值相符。结论实验所用不同来源弓形虫分离株的GRA1基因扩增片段基本相同，且3种限制性内切酶酶切图诺基本一致，表明这3个分离株的GRA1基因高度保守。

Aim Comparison of amplification of GRA1 gene in vitro from different geographical and heterologousToxoplasma gondii isolates RH (human), ZS_2(human), CN (pig) in study. Methods Using polymerase chain reaction (PCR)and restriction analysis techniques,the specific fragments of GRA1 gene were amplified from the genomic DNA of Toxoplasmagondii RH,ZS_2,CN isolates. The fragments of GRA1 gene were digested by Hind F,HPa Ⅱ,Taq Ⅰ in the restriction endonu-clease reaction,which had been identified in agarose gel electrophorosis. Results The specific fragments of GRA1 gene weresuccessfully amplified from the genomic DNA of RH isolate,CN isolate,ZS_2 isolate of Toxoplasma gondii. The fragments ofPCR amplification digested with Hind Ⅲ,Hpa Ⅱ,Taq Ⅰ were not significantly different. Conclusion It demonstracted highlyconservation among heterologous Toxoplasma isolates GRAl genes.