不同分离株弓形虫hsp70基因体外扩增及其酶切分析

目的　不同分离株弓形虫hsp70基因的比较 ;运用限制性内切酶分析 (PRA)建立一种简单的虫株鉴定方法。 方法　采用PCR技术扩增hsp70基因 ;采用 3种限制性内切酶Bsp12 86Ⅰ、BstXⅠ、MboⅠ分别对该基因进行酶切比较。 结果　从 5种不同分离株的弓形虫基因组DNA中扩增出相似的约 2 0 2 2bp的完整HSP70基因阅读框序列 ;Bsp12 86Ⅰ酶切 5株弓形虫有 2组不同酶谱 ,而BstXⅠ、MboⅠ酶切的片段各株间没有差异。结论　 5种不同分离株弓形虫的hsp70基因片段大小基本相同 ;但酶切图谱有差异 ,这可以为虫株鉴定和毒力研究提供参考。

Amplification of the hsp70 genes among different geographical strains of isolates of Toxoplasma gondii and its restriction fragment length analysis

To study the hsp70 genes among different geographical strains of isolates of Toxoplasma gondii,and to establish a simple technique with PCR restriction enzyme analysis(PRA) for the rapid identification of isolates,the hsp70 gene was amplified by PCR and followed by restriction enzyme analysis with the gene digested by Bsp1286 Ⅰ,BstXⅠ and MboⅠ.It was found that similar hsp70 gene ORF with approximate 2022 bp fragment was successfully amplified from the genomes of 5 different geographical isolates,in which two different types of pattern of endonuclease digestion in the hsp70 gene digested with Bsp1286 I were found,but there was no any difference among those digested by BstX Ⅰ or Mbo.It concludes that the size of hsp70 gene fragments of 5 strains of Toxoplasma gondii isolated from different geographical areas are almost the same,but they show difference in the endonuclease digestion pattern.This properties could provide references for the identification and virulence characterization of Toxoplasma gondii.