PROPER EXPRESSION OF MYOSIN GENES IN TRANSGENIC NEMATODES
PROPER EXPRESSION OF MYOSIN GENES IN TRANSGENIC NEMATODES
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DOI：
10.1002/j.1460-2075.1989.tb08506.x
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发表时间：
1989-11-01
期刊：
影响因子：
11.4
通讯作者：
WATERSTON, RH
中科院分区：
文献类型：
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作者：
FIRE, A;WATERSTON, RH
秀丽隐杆线虫有四个编码骨骼肌球蛋白重链亚型的基因。我们已将其中两个基因 myo-3 和 unc-54 的克隆以低拷贝数重新引入秀丽隐杆线虫种系中。根据两个遗传标准，所得基因座表现为基因的功能副本：（i）它们可以导致携带失活 myo-3 或 unc-54 突变的菌株的表型拯救，以及（ii）它们存在于具有内源肌球蛋白基因座野生型副本的菌株中，具有与复制内源基因座类似的遗传后果。重新引入的基因的功能水平接近于内源基因座的水平。针对不同亚型的特异性单克隆抗体已用于定位表达的蛋白质。重新引入的基因在与内源基因完全相同的细胞类型中表达，并且产生的肌球蛋白产物组装成与野生型中一样的丝状结构。出乎意料的是，我们在这项工作过程中发现，非常高的 unc-54 基因拷贝数会导致肌肉结构的破坏，这可能是由于蛋白质产物的过度表达造成的。
Caenorhabditis elegans has four genes which encode skeletal myosin heavy chain isoforms. We have reintroduced clones of two of these genes, myo-3 and unc-54 at low copy number into the germline of C. elegans. The resulting loci behave as functional copies of the genes by two genetic criteria: (i) they can result in phenotypic rescue of strains carrying inactivating myo-3 or unc-54 mutations, and (ii) their presence in strains with wild-type copies of the endogenous myosin loci has genetic consequences similar to duplicating the endogenous loci. The re-introduced genes function at a level close to that of the endogenous loci. Monoclonal antibodies specific for the different isoforms have been used to localize the expressed proteins. The re-introduced genes express in precisely the same cell types as the endogenous genes, and the myosin products produced assemble into filament structures as in wild-type. Unexpectedly, we have found in the course of this work that very high copy numbers of the unc-54 gene lead to a disruption of muscle structure which may result from overexpression of the protein product.