Gomes M L, Romanha A J, Gonçalves A M, Chiari E
Universidade Estadual de Maringá, Depto. de Análises Clínicas, Maringá, Brasil.
Mem Inst Oswaldo Cruz. 1991 Oct-Dec;86(4):379-85. doi: 10.1590/s0074-02761991000400001.
Four Trypanosoma cruzi strains from zymodemes A, B, C and D were successively cloned on BHI-LIT-agar-blood (BLAB). Twenty clones from the first generation (F1), 10 from the second (F2) and 4 from the third (F3) from the strains A138, B147 and C231 were isolated. The D150 strain provided 29 F1 and 23 F2 clones. The strains and clones had their isoenzyme and k-DNA patterns determined. The clones from A138, B147 and C231 strains presented isoenzyme and k-DNA patterns identical between themselves and their respective parental strains. Therefore showing the homogeneity and stability of isoenzyme and k-DNA patterns after successive cloning. The D150 strain from zymodeme D (ZD) showed heterogeneity. Twenty-eight out of 29 clones of the first generation were of zymodeme A and only one was of zymodeme C, confirming previous reports that ZD strains consisted of ZA and ZC parasite populations. The only D150 strain clone of zymodeme C showed a k-DNA pattern identical to its parental strain. The remaining clones although similar among themselves were different from the parental strain. Thus the T. cruzi strains had either homonogeneus or heterogeneous populations. The clones produced by successive cloning provided genetically homogeneous populations. Their experimental use will make future results more reliable and reproducible.
从酶谱型A、B、C和D中选取了4株克氏锥虫菌株，先后在脑心浸液-肝浸液-琼脂-血液（BLAB）培养基上进行克隆。从A138、B147和C231菌株的第一代（F1）中分离出20个克隆，第二代（F2）中分离出10个克隆，第三代（F3）中分离出4个克隆。D150菌株产生了29个F1克隆和23个F2克隆。对这些菌株和克隆进行了同工酶和k-DNA图谱测定。A138、B147和C231菌株的克隆呈现出与各自亲本菌株相同的同工酶和k-DNA图谱。因此表明连续克隆后同工酶和k-DNA图谱具有同质性和稳定性。来自酶谱型D（ZD）的D150菌株表现出异质性。第一代的29个克隆中有28个属于酶谱型A，只有1个属于酶谱型C，这证实了之前关于ZD菌株由ZA和ZC寄生虫群体组成的报道。酶谱型C的唯一D150菌株克隆呈现出与其亲本菌株相同的k-DNA图谱。其余克隆虽然彼此相似，但与亲本菌株不同。因此，克氏锥虫菌株具有同质性或异质性群体。连续克隆产生的克隆提供了基因同质的群体。它们在实验中的应用将使未来的结果更可靠且可重复。
