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[通过荧光原位杂交对转化基因进行定位]

[Mapping of the transforming gene by fluorescence in situ hybridization].

作者信息

He H

机构信息

Tumor Research Institute, Hunan Medical University, Changsha.

出版信息

Zhonghua Yi Xue Za Zhi. 1993 Jun;73(6):338-40, 380.

PMID:8258102
Abstract

A transforming gene of human nasopharyngeal carcinoma was cloned from Chinese nasopharyngeal carcinoma cell line CNE2. This gene was different from a number of known oncogenes as was reported previously. The NPC transforming gene was 16kb in size, and the Alu negative 2.8kb fragment was cloned into pSP70 plasmid. Linearized Tx pSP70 recombinant plasmid was nick translated in the presence of biotin-11-dUTP, incubated with an excess of sonicated human DNA, and used as a probe for fluorescence in situ hybridization. Signals of hybridization were detected by successive treatments with FITC-labeled avidin. 57% of grains were located on chromosome 8; 51.8% of them were mapped to the 8q23-->8q24.1 region of chromosome 8's. The Tx gene, therefore, could be assigned to the band 8q23-->8q24.1.

摘要

从中国鼻咽癌细胞系CNE2中克隆出一种人类鼻咽癌转化基因。该基因与先前报道的许多已知癌基因不同。鼻咽癌转化基因大小为16kb,将Alu阴性的2.8kb片段克隆到pSP70质粒中。线性化的Tx pSP70重组质粒在生物素-11-dUTP存在的情况下进行缺口平移,与过量的超声破碎的人类DNA一起温育,并用作荧光原位杂交的探针。通过用异硫氰酸荧光素(FITC)标记的抗生物素蛋白连续处理来检测杂交信号。57%的颗粒位于8号染色体上;其中51.8%定位于8号染色体的8q23→8q24.1区域。因此,Tx基因可定位于8q23→8q24.1带。

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