FLUORESCENCE INSITU HYBRIDIZATION WITH ALU AND L1 POLYMERASE CHAIN-REACTION PROBES FOR RAPID CHARACTERIZATION OF HUMAN-CHROMOSOMES IN HYBRID CELL-LINES
FLUORESCENCE INSITU HYBRIDIZATION WITH ALU AND L1 POLYMERASE CHAIN-REACTION PROBES FOR RAPID CHARACTERIZATION OF HUMAN-CHROMOSOMES IN HYBRID CELL-LINES
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DOI：
10.1073/pnas.87.17.6634
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发表时间：
1990-09-01
影响因子：
11.1
通讯作者：
WARD, DC
中科院分区：
文献类型：
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作者：
LICHTER, P;LEDBETTER, SA;WARD, DC
人类-啮齿动物杂交细胞系的人类DNA含量已被分析，使用不同的DNA探针组，从杂交衍生，原位杂交到正常的人类中期染色体扩散。将总基因组杂交DNA与在人类序列中高度富集的杂交DNA探针组进行比较。后一种探针是通过聚合酶链反应（PCR），利用寡核苷酸引物对穿插重复序列Alu和L1的人类特异性子序列进行扩增而获得的。在杂交系中以前未识别的染色体物质以快速和精确的方式进行了表征。结果表明，通过比较Alu-PCR产物与L1-PCR产物或基因组杂交DNA获得的数据，可以快速评估杂交系的完整人类补体。这种使用穿插重复序列pcr产物的方法简单快速，也提供了一种通过原位杂交生成复杂DNA探针集的替代方法，用于对整个染色体或亚染色体区域进行特定的描绘。
Human-rodent hybrid cell lines have been analyzed with regard to their human DNA content by using various DNA probe sets, derived from the hybrids, for in situ hybridization to normal human metaphase chromosome spreads. Total genomic hybrid DNA was compared with probe sets of hybrid DNA that were highly enriched in human sequences. The latter probes were obtrained by amplification through the polymerase chain reaction (PCR) using oligonucleotide primers directed to human specific subsequences of the interspersed repetititve sequences Alu and L1. Previously unidentified chromosomal material within hybrid lines was characterized with speed and precision. It is demonstrated that the complete human complement of hybrid lines can be rapidly assessed by comparing the data obtained with the Alu-PCR products with the results from the L1-PCR products or from the genomic hybrid DNA. This approach using interspersed repetitive sequence-PCR products is simple and fast and also provides an alternative way of generating complex DNA probe sets for the specific delineation of entire chromosomes or subchromosomal regions by in situ hybridization.