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大核移植诱导四膜虫刺丝泡表型转变

Trichocyst phenotype transformation induced by macronuclear transplantation in Paramecium tetraurelia.

作者信息

Aufderheide K J

出版信息

Exp Cell Res. 1985 Jan;156(1):282-6. doi: 10.1016/0014-4827(85)90283-6.

Abstract

A portion of the macronucleus of wild-type cells of Paramecium tetraurelia was removed and was injected into cells homozygous for the ftA mutation. The ftA mutants make defective trichocysts and are unable to perform normal trichocyst exocytosis. After injection, approx. 30% of the surviving cells show a phenotype shift from mutant to wild-type. This shift is stable during subsequent vegetative growth until clonal death. If, however, the hybrid cell lines are brought to autogamy (which discards the existing macronucleus and forms a new one from sexual products derived from a micronucleus), then the lines revert to the ftA phenotype. Since micronuclei were not transplanted, the phenotypic reversion after autogamy is to be expected, and demonstrates that the transformation affects the macronucleus only. A second series of injections involved transfer of a portion of the macronucleus from cells homozygous for the trichocyst ptA mutation into ftA host cells. These two mutations are genetically complementary, so the injection should be genetically equivalent to forming a double heterozygote. Approx. 20% of the injection survivors shift to wild-type. This shift is also vegetatively stable unless autogamy occurs; after autogamy, reversion to the ftA phenotype is seen. These results show that a portion of a macronucleus can be successfully transplanted from one cell to another and that, in the host cytoplasmic environment, normal gene expression and replication of a transplanted macronucleus does occur. The technique of macronuclear transplantation is significant to studies of the macronuclear contribution to clonal aging, and to studies on genetic control over trichocyst development.

摘要

切除了四膜虫野生型细胞的一部分大核,并将其注射到ftA突变纯合的细胞中。ftA突变体形成有缺陷的刺丝泡,并且无法进行正常的刺丝泡胞吐作用。注射后,约30%存活的细胞表现出从突变型到野生型的表型转变。这种转变在随后的营养生长过程中一直稳定,直到克隆死亡。然而,如果将杂种细胞系进行自体受精(这会丢弃现有的大核,并从源自小核的有性产物中形成一个新的大核),那么这些细胞系会恢复到ftA表型。由于没有移植小核,自体受精后的表型逆转是可以预期的,这表明这种转变仅影响大核。第二组注射涉及将来自刺丝泡ptA突变纯合细胞的一部分大核转移到ftA宿主细胞中。这两个突变在遗传上是互补的,因此这种注射在遗传上相当于形成一个双杂合子。约20%注射存活的细胞转变为野生型。这种转变在营养生长过程中也是稳定的,除非发生自体受精;自体受精后,会出现恢复到ftA表型的情况。这些结果表明,一部分大核可以成功地从一个细胞移植到另一个细胞,并且在宿主细胞质环境中,移植的大核确实会发生正常的基因表达和复制。大核移植技术对于研究大核对克隆衰老的贡献以及对刺丝泡发育的遗传控制研究具有重要意义。

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