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禽骨髓细胞瘤病毒(MC29)的转化序列。

The transforming sequences of avian myelocytomatosis virus (MC29).

作者信息

Lautenberger J A, Schulz R A, Garon C F, Tsichlis P N, Spyropoulos D D, Pry T W, Rushlow K E, Papas T S

出版信息

J Supramol Struct Cell Biochem. 1981;16(2):193-207. doi: 10.1002/jsscb.1981.380160209.

Abstract

Avian myelocytomatosis virus (MC29), a defective acute leukemia virus, has a broad oncogenic spectrum in vivo, and transforms fibroblasts and hematopoietic target cells in vitro, We have used recombinant DNA technology to isolate and characterize the sequences that are essential in the transformation process. Integrated MC29 proviral DNA was isolated from a library of recombinant phage containing DNA from the MC29-transformed nonproducer quail cell line Q5. The cloned DNA was analyzed by Southern blotting of restriction endonuclease digests and by electron microscopic visualization of R-loops formed between the cloned DNA and MC29 or helper virus RNA. It was found that the 9.2 kb cloned DNA insert contains approximately 4 kb of viral sequences and 5.2 kb of quail cellular sequences. The viral sequences contain all of the MC29 specific sequences and 5' helper related sequences as well as part of the envelope region. The size of the cloned EcoRI fragment is the same as that of the major band in EcoRI-cleaved Q5 DNA that hybridizes to viral sequences. Transfection of the cloned DNA into NIH 3T3 cells revealed that the MC29-specific sequences are functional in that they induce foci of transformed cells with high efficiency.

摘要

禽骨髓细胞瘤病毒(MC29)是一种缺陷型急性白血病病毒,在体内具有广泛的致癌谱,在体外可转化成纤维细胞和造血靶细胞。我们利用重组DNA技术分离并鉴定了转化过程中必需的序列。整合的MC29前病毒DNA是从一个重组噬菌体文库中分离得到的,该文库包含来自MC29转化的非生产性鹌鹑细胞系Q5的DNA。通过对限制性内切酶消化产物进行Southern印迹分析以及通过电子显微镜观察克隆DNA与MC29或辅助病毒RNA之间形成的R环,对克隆的DNA进行了分析。结果发现,9.2 kb的克隆DNA插入片段包含约4 kb的病毒序列和5.2 kb的鹌鹑细胞序列。病毒序列包含所有MC29特异性序列、5'辅助相关序列以及包膜区域的一部分。克隆的EcoRI片段的大小与EcoRI切割的Q5 DNA中与病毒序列杂交的主要条带的大小相同。将克隆的DNA转染到NIH 3T3细胞中发现,MC29特异性序列具有功能,因为它们能高效诱导转化细胞灶。

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