通过酚乳液竞争性DNA重退火快速克隆人神经母细胞瘤细胞系的多个扩增核苷酸序列

Rapid cloning of multiple amplified nucleotide sequences from human neuroblastoma cell lines by phenol emulsion competitive DNA reassociation.

作者信息

Shiloh Y, Rose E, Colletti-Feener C, Korf B, Kunkel L M, Latt S A

出版信息

Gene. 1987;51(1):53-9. doi: 10.1016/0378-1119(87)90473-2.

Abstract

A protocol for the rapid cloning of many DNA fragments from an amplified genomic region is described. The procedure is based on a modification of the phenol-emulsion reassociation technique (PERT) previously used to clone DNA fragments missing from a chromosomal deletion [Kunkel et al., Proc. Natl. Acad. Sci. USA 82 (1985) 4778-4782]. The procedure was used to construct recombinant libraries in the plasmid pBR322 which were highly enriched for amplified sequences from two neuroblastoma cell lines, CHP-126 and IMR-32. Many new amplified DNA fragments were isolated from these libraries, indicating that the PERT methodology should be of general use in isolating amplified DNA from other cell lines and tumors.

摘要

本文描述了一种从扩增的基因组区域快速克隆多个DNA片段的方法。该方法基于对酚乳液复性技术（PERT）的改进，此前该技术用于克隆染色体缺失中缺失的DNA片段[Kunkel等人，《美国国家科学院院刊》82 (1985) 4778 - 4782]。该方法用于在质粒pBR322中构建重组文库，这些文库高度富集了来自两种神经母细胞瘤细胞系CHP - 126和IMR - 32的扩增序列。从这些文库中分离出许多新的扩增DNA片段，表明PERT方法在从其他细胞系和肿瘤中分离扩增DNA方面应具有普遍用途。