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从人肝细胞癌中克隆一个癌基因

Molecular cloning of an oncogene from a human hepatocellular carcinoma.

作者信息

Ochiya T, Fujiyama A, Fukushige S, Hatada I, Matsubara K

出版信息

Proc Natl Acad Sci U S A. 1986 Jul;83(14):4993-7. doi: 10.1073/pnas.83.14.4993.

Abstract

A transforming DNA, named lca (for liver cancer), was obtained from a primary human hepatocellular carcinoma (HCC) in transformation assays using NIH 3T3 cells and a calcium phosphate coprecipitation method. High molecular weight DNA obtained from the HCC tissue was employed for this purpose. This transforming DNA had a linkage to the Alu sequence and was cloned in lambda phage for further studies. Restriction enzyme analyses showed that the minimal size of the lca transforming DNA is about 10 kilobase pairs and that its cleavage profiles are different from those of any one of the previously reported human transforming genes or retroviral oncogenes. No cross-hybridization was observed between these genes and the lca DNA. Southern blot analyses of DNAs from flow-sorted human chromosomes and human-mouse somatic cell hybrids indicated that the lca DNA is located on human chromosome 2. An independently obtained transforming DNA from another HCC exhibited identical restriction enzyme cleavage profiles. Thus, lca DNA is likely to represent a commonly encountered transforming DNA in HCC.

摘要

一种名为lca(肝癌之意)的转化DNA,是通过使用NIH 3T3细胞和磷酸钙共沉淀法的转化试验,从原发性人类肝细胞癌(HCC)中获得的。为此采用了从HCC组织中获取的高分子量DNA。这种转化DNA与Alu序列有关联,并被克隆到λ噬菌体中用于进一步研究。限制性内切酶分析表明,lca转化DNA的最小大小约为10千碱基对,其切割图谱与先前报道的任何一种人类转化基因或逆转录病毒癌基因的切割图谱都不同。这些基因与lca DNA之间未观察到交叉杂交。对来自流式分选的人类染色体和人 - 鼠体细胞杂种的DNA进行的Southern印迹分析表明,lca DNA位于人类2号染色体上。从另一个HCC独立获得的转化DNA表现出相同的限制性内切酶切割图谱。因此,lca DNA可能代表HCC中常见的转化DNA。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15e9/323875/6d7d2fbca96f/pnas00318-0029-a.jpg

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