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源自勒克肾癌细胞核克隆的胚胎和蝌蚪中疱疹病毒DNA的命运

Fate of herpesvirus DNA in embryos and tadpoles cloned from Lucké renal carcinoma nuclei.

作者信息

Carlson D L, Sauerbier W, Rollins-Smith L A, McKinnell R G

机构信息

Department of Genetics and Cell Biology, University of Minnesota, Saint Paul 55108.

出版信息

J Comp Pathol. 1994 Aug;111(2):197-204. doi: 10.1016/s0021-9975(05)80051-9.

Abstract

The Lucké renal carcinoma of the northern leopard frog, Rana pipiens, has a herpesvirus aetiology. Lucké tumour nuclei inserted into enucleated frogs' eggs produce development to the swimming tadpole stage. Tissue from these tumour nuclear transplant animals can be induced to survive and differentiate further by allografting to normal tadpoles. We wished to ascertain whether the aetiological agent, the Lucké tumour herpesvirus (LTV), persists in the animals produced by tumour nuclear transplantation. The polymerase chain reaction was used to amplify a 1.2 kbp Hind III restriction fragment of LTV DNA in whole animal homogenates prepared from tumour nuclear transplant tadpoles and normal tadpoles fertilized in vitro. The LTV fragment was not present in the majority (31 of 34) of the cloned animals derived from tumour nuclei, nor was it present in any of five normal tadpoles. Either the 1.2 kbp fragment of LTV DNA was eliminated from most of the cloned animals during the massive reprogramming of the neoplastic genome initiated by insertion of the tumour nuclei into egg cytoplasm, or the nuclei selected for transplantation were primarily those lacking this fragment of LTV DNA. The limited development of the tumour nuclear tadpoles was probably not due to the presence of these viral sequences, but rather reflected the limited plasticity of the tumour cell genome as assayed by nuclear transplantation. Failure to detect the 1.2 kbp fragment of LTV DNA in the majority of mitotic progeny of the Lucké tumour genome does not imply that other parts of the viral genome do not persist.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

北美豹蛙(Rana pipiens)的勒克肾腺癌由一种疱疹病毒引起。将勒克肿瘤细胞核植入去核蛙卵后可发育至游动蝌蚪阶段。这些肿瘤核移植动物的组织通过移植到正常蝌蚪体内可被诱导进一步存活和分化。我们希望确定病因病原体,即勒克肿瘤疱疹病毒(LTV),是否在肿瘤核移植产生的动物体内持续存在。利用聚合酶链反应,在由肿瘤核移植蝌蚪和体外受精的正常蝌蚪制备的全动物匀浆中扩增LTV DNA的1.2 kbp Hind III限制性片段。在大多数(34只中的31只)源自肿瘤核的克隆动物中未发现LTV片段,五只正常蝌蚪中也均未发现。要么在将肿瘤核插入卵细胞质引发肿瘤基因组大规模重编程过程中,大多数克隆动物体内的LTV DNA的1.2 kbp片段被消除,要么用于移植的细胞核主要是那些缺乏LTV DNA该片段的细胞核。肿瘤核蝌蚪发育受限可能并非由于这些病毒序列的存在,而是反映了通过核移植检测到的肿瘤细胞基因组可塑性有限。在勒克肿瘤基因组的大多数有丝分裂后代中未检测到LTV DNA的1.2 kbp片段,并不意味着病毒基因组的其他部分不存在。(摘要截短至250字)

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