CLONING AND SEQUENCING OF CDNA-ENCODING HUMAN DNA TOPOISOMERASE-II AND LOCALIZATION OF THE GENE TO CHROMOSOME REGION 17Q21-22
CLONING AND SEQUENCING OF CDNA-ENCODING HUMAN DNA TOPOISOMERASE-II AND LOCALIZATION OF THE GENE TO CHROMOSOME REGION 17Q21-22
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DOI：
10.1073/pnas.85.19.7177
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发表时间：
1988-10-01
影响因子：
11.1
通讯作者：
WANG, JC
中科院分区：
文献类型：
--
作者：
TSAIPFLUGFELDER, M;LIU, LF;WANG, JC
两个重叠的cDNA克隆编码人DNA拓扑异构酶II的两个独立的方法进行了鉴定。在一个中，噬菌体中的人cDNA文库。通过与编码在酵母和果蝇DNA拓扑异构酶II中发现的一段七个氨基酸的混合寡核苷酸探针杂交来筛选;在另一个中，针对抗原决定物的表达来筛选在一个不同的人cDNA文库中的人gt 11表达载体，所述抗原决定物被特异于人DNA拓扑异构酶II的兔抗体识别。从这些和几个额外的克隆中确定了人DNA拓扑异构酶II基因的整个编码序列，通过使用克隆的人TOP 2基因序列作为探针进行鉴定。克隆序列与mRNA和基因组DNA之间的杂交表明，人类酶由单拷贝基因编码。通过将克隆片段与中期染色体进行原位杂交，并通过与一组小鼠-人杂交细胞系进行杂交分析，将该基因的位置定位于染色体17 q21 -22，每个杂交细胞系保留一个人类染色体子集。
Two overlapping cDNA clones encoding human DNA topoisomerase II were identified by two independent methods. In one, a human cDNA library in phage .lambda. was screened by hybridization with a mixed oligonucleotide probe encoding a stretch of seven amino acids found in yeast and Drosophila DNA topoisomerase II; in the other, a different human cDNA library in a .lambda.gt11 expression vector was screened for the expression of antigenic deteminants that are recognized by rabbit antibodies specific to human DNA topoisomerase II. The entire coding sequences of the human DNA topoisomerase II gene were determined from these and several additional clones, identified through the use of the cloned human TOP2 gene sequences as probes. Hybridization between the cloned sequences and mRNA and genomic DNA indicates that the human enzyme is encoded by a single-copy gene. The location of the gene was mapped to chromosome 17q21-22 by in situ hybridization of a cloned fragment to metaphase chromosomes and by hybridization analysis with a panel of mouse-human hybrid cell lines, each retaining a subset of human chromosomes.