Johnson A M, Illana S, Dubey J P, Dame J B
Exp Parasitol. 1987 Jun;63(3):272-8. doi: 10.1016/0014-4894(87)90173-1.
A mung bean nuclease genomic library of purified DNA from tachyzoites of the RH strain of Toxoplasma gondii was prepared in the bacteriophage lambda gtll and recombinants containing rRNA gene fragments were detected by hybridization with radiolabeled total RNA from the closely related coccidian Eimeria acervulina. Ten recombinants were chosen at random, and five of these were investigated further using probes for the genes of the large and small rRNA of Plasmodium berghei. An insert (called TG4) that hybridized only to the 3' end of the large rRNA coding region of P. berghei and an insert (called TG18) that hybridized only to the small rRNA coding region of P. berghei were purified by electrophoresis in low melting point agarose. Radiolabeled E. acervulina total RNA, TG4, and TG18, were then used to compare the sizes of the large and small rRNA gene fragments after DNA extracted from three strains of T. gondii, and the type strain of the closely related coccidian Hammondia hammondi were cut by one of a series of 10 restriction endonucleases. The patterns obtained for the three T. gondii isolates were identical to those obtained for H. hammondi, for each enzyme tested. In addition, the guanine plus cytosine (G + C) content of H. hammondi DNA was found to be almost identical to that obtained previously for T. gondii DNA.
从刚地弓形虫RH株速殖子中提取纯化DNA,构建了绿豆核酸酶基因组文库,该文库以噬菌体λgtll为载体。通过与来自密切相关的球虫艾美耳球虫的放射性标记总RNA杂交,检测含有rRNA基因片段的重组体。随机选择10个重组体,其中5个使用伯氏疟原虫大、小rRNA基因探针进一步研究。通过在低熔点琼脂糖中电泳纯化出仅与伯氏疟原虫大rRNA编码区3'端杂交的插入片段(称为TG4)和仅与伯氏疟原虫小rRNA编码区杂交的插入片段(称为TG18)。然后,用放射性标记的艾美耳球虫总RNA、TG4和TG18,比较从三株刚地弓形虫以及密切相关的球虫哈蒙德球虫模式株中提取的DNA,用一系列10种限制性内切酶之一切割后大、小rRNA基因片段的大小。对于所测试的每种酶,三株刚地弓形虫分离株获得的图谱与哈蒙德球虫获得的图谱相同。此外,发现哈蒙德球虫DNA的鸟嘌呤加胞嘧啶(G + C)含量与先前获得的刚地弓形虫DNA的含量几乎相同。