HIGH-EFFICIENCY CLONAL GROWTH OF BLOOD-STREAM-FORM AND INSECT-FORM TRYPANOSOMA-BRUCEI ON AGAROSE PLATES
HIGH-EFFICIENCY CLONAL GROWTH OF BLOOD-STREAM-FORM AND INSECT-FORM TRYPANOSOMA-BRUCEI ON AGAROSE PLATES
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DOI：
10.1073/pnas.89.18.8818
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发表时间：
1992-09-15
影响因子：
11.1
通讯作者：
CROSS, GAM
中科院分区：
文献类型：
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作者：
CARRUTHERS, VB;CROSS, GAM
本报告描述了一种在琼脂糖平板上生长血流型和顺环型布氏锥虫菌落的方法。前环菌落需要2周的时间才能形成，在补充有20%（体积/体积）条件培养基的SDM-79/0.65%琼脂糖上以约17%的平板效率生长。使血流形式适应于在液体HMI-9培养基中的体外生长，然后涂布在HMI-9/0.65%琼脂糖平板上，在那里它们在3-5天内生长成可见的菌落。平板接种效率为3 - 80%，取决于锥虫变体和实验。经证明，菌落是单细胞生长的结果，成熟时含有约10（6）个细胞。将菌落转移到过滤器中，并探测多拷贝和单拷贝基因。这种方法结合经典和反向遗传学方法研究锥虫的潜在用途进行了讨论。
This report describes a method for growing both bloodstream- and procyclic-form Trypanosoma brucei as colonies on agarose plates. Procyclic colonies, which took 2 weeks to develop, grew with approximately 17% plating efficiency on SDM-79/0.65% agarose supplemented with 20% (vol/vol) conditioned medium. Bloodstream forms were adapted to in vitro growth in liquid HMI-9 medium and then spread on HMI-9/0.65% agarose plates, where they grew to visible colonies in 3-5 days. Plating efficiencies were from 3 to 80%, depending upon the trypanosome variant and experiment. Colonies were proven to be the result of growth from a single cell and contained approximately 10(6) cells at maturity. Colonies were transferred to filters and probed for multicopy and single-copy genes. Potential uses of this method in conjunction with classical and reverse genetic approaches to studying trypanosomes are discussed.