Tanriverdi Sultan, Blain J Craig, Deng Bingbing, Ferdig Michael T, Widmer Giovanni
Tufts Cummings School of Veterinary Medicine, Division of Infectious Diseases, North Grafton, MA, USA.
Mol Microbiol. 2007 Mar;63(5):1432-9. doi: 10.1111/j.1365-2958.2007.05594.x.
Recombinant progeny lines of Cryptosporidium parvum were generated by coinfecting immunosuppressed mice with two genetically distinct isolates of C. parvum. Progeny lines were obtained from a cross of parental lines MD x TU114 through targeted propagation in mice of progeny oocysts originating from populations lacking one parental allele at one or more loci. For each infection lineage this process was repeated until only a single allele remained for each marker, indicating that the progeny line was clonal. To study genetic recombination, 16 progeny clones were genotyped at 40 loci located on each of the eight chromosomes. The inheritance of parental alleles was significantly skewed towards the more virulent parent isolate MD. A contiguous 476 kb segment of chromosome V displayed MD allele in all progeny recovered, while MD and TU114 alleles were detected at other loci throughout the genome. The absence of alleles from one parental isolate in this chromosomal region may indicate phenotypic selection for the MD allele during the generation of these lines. A range for the meiotic crossover frequency was determined on the basis of 40 markers and the number of meioses estimated to have taken place during the crossing experiment. C. parvum exhibits a high rate of recombination commensurate with other Apicomplexa.
通过用两种基因不同的微小隐孢子虫分离株共同感染免疫抑制小鼠,产生了微小隐孢子虫的重组后代品系。通过在小鼠中对源自一个或多个位点缺乏一个亲本等位基因的群体的后代卵囊进行靶向繁殖,从亲本品系MD×TU114的杂交中获得了后代品系。对于每个感染谱系,重复这个过程,直到每个标记只剩下一个等位基因,这表明后代品系是克隆的。为了研究基因重组,对位于八条染色体上的40个位点的16个后代克隆进行了基因分型。亲本等位基因的遗传明显偏向于毒性更强的亲本分离株MD。在回收的所有后代中,第五条染色体上一个连续的476 kb片段显示为MD等位基因,而在整个基因组的其他位点检测到MD和TU114等位基因。在这个染色体区域中没有一个亲本分离株的等位基因,这可能表明在这些品系的产生过程中对MD等位基因进行了表型选择。根据40个标记以及估计在杂交实验中发生的减数分裂次数,确定了减数分裂交换频率的范围。微小隐孢子虫表现出与其他顶复门生物相当的高重组率。