Novel strategies to clone identical and distinct DNA sequences for several complex genomes |
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Authors: | E R Zabarovsky |
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Institution: | (1) Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 117984 Moscow, Russia;(2) Microbiology & Tumor Biology Center and Center for Genomics Research, Karolinska Institute, Box 280, S-171 77 Stockholm, Sweden |
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Abstract: | In this minireview I briefly describe the new methods suggested for cloning sequences identical by descent, homo-or hemizygously
deleted, amplified or polymorphic, and compare them with the most efficient techniques developed earlier. The new methods
include cloning of identical sequences (CIS), cloning of polymorphic sequences (COP), and cloning of deleted sequences (CODE).
Although these methods are based on the same combination of biochemical techniques, their aims are different. These methods
are fully complementary, and they may be combined to analyze a given object. If one aims to clone a disease gene responsible
for familial cancer syndrome, these methods may be applied as follows. CIS can be used to identify the sequences identical
by descent comparing the DNA obtained from affected or unaffected family members. COP can be used to find sequences that are
different between affected and unaffected members, and CODE would be useful to compare tumor and normal (control) samples
to isolate, deleted sequences (putative candidate tumor suppressor genes) and amplified sequences (putative oncogenes). The
COP and CODE procedures can be applied to analyze the CpG islands, thus allowing direct candidate gene identification. |
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Keywords: | gene deletion gene amplification human genome molecular cloning subtractive hybridization polymorphism chromosome mapping genome mismatch scanning |
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