用于分子杂交的染色体。使用快速滤膜固定法对重复基因和单拷贝基因进行定位。

Chromosomes for molecular hybridization. Assignment of repetitive and single copy genes using a rapid filter-fixation method.

作者信息

Langer G, Blin N, Stoehr M

出版信息

Histochemistry. 1984;80(5):469-73. doi: 10.1007/BF00495436.

Abstract

Specific recombinant DNA sequences (5S rRNA, B1, albumin) were assigned to flow sorted chromosomes of the Chinese hamster cell line CHV79. For this purpose, a rapid protocol was developed using filterbound chromosomal DNA and probing with various nucleic acids, that allows sequence identification in chromosomes. A flow histogram and a flow karyogram of the CHV79 cell line were established by flow analysis in order to calculate the amount of DNA per CHV79 cell and their chromosomes. Subsequently, metaphase chromosomes or chromosomal groups were fractionated by electronic sorting and a defined number of chromosomes was directly bound to nitrocellulose filters for sequence homology analysis by a dot blot hybridization procedure. This procedure not only allows the assigning of specific DNA sequences to particular chromosomes, it is also applicable to studies of changes in karyotypes, for example translocations of given sequences.

摘要

特定的重组DNA序列（5S rRNA、B1、白蛋白）被定位到中国仓鼠细胞系CHV79经流式分选的染色体上。为此，开发了一种快速方案，该方案使用滤膜结合的染色体DNA并用各种核酸进行探针杂交，从而能够在染色体中鉴定序列。通过流式分析建立了CHV79细胞系的流式直方图和流式核型图，以计算每个CHV79细胞及其染色体的DNA含量。随后，通过电子分选对中期染色体或染色体组进行分离，并将一定数量的染色体直接结合到硝酸纤维素滤膜上，通过斑点杂交程序进行序列同源性分析。该程序不仅能够将特定的DNA序列定位到特定的染色体上，还适用于核型变化的研究，例如特定序列的易位。