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© 1995 Oxford University Press

OTHER

Isolation of chromosome-specific genes by reciprocal probing of arrayed cDNA and cosmid libraries

Cheng Chi Lee1,+,*, Ali Yazdani1,{ddagger}, Manfred Wehnert1,{ddagger},§, ZhaoYang Zhao2, Elizabeth A. Lindsay1, Jennifer Bailey2, Mary I. Coolbaugh2, Lisa Couch1, Mengli Xiong1, A.Craig Chinault1, Antonio Baldini1 and C.Thomas Caskey1,2,¶

1Department of Molecular and Human Genetics, Baylor College of Medicine Houston, TX77030 2Howard Hughes Medical Institute, Baylor College of Medicine Houston, TX77030, USA {ddagger}These authors made equal contribution to the research +Clayton Foundation for Research §Currently at Institute for Medical Genetics, University of Greifswald Greifswald, Germany Currently at MERCK Research Labs Summeytown, West Point, PA 19486, USA

*To whom correspondence should be addressed

Received March 9, 1995; Revised May 10, 1995; Accepted May 10, 1995

We have identified and mapped 61 novel and previously described chromosome 17 and X genes, using a human placental cDNA library. These genes were isolated using a gene identification and mapping strategy based on reciprocal probing of arrayed chromosome specific cosmid and cDNA libraries. This strategy scans gridded cosmids for nuclear genes and isolates the expressed sequence by a cosmid to cDNA filter hybridization. Inherent to this approach is the advantage of identifying the corresponding genomic cosmid clone of a particular cDNA. The genomic and cDNA reagents can be used for fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR) based mapping to resolve map positions of cDNAs belonging to gene families and those associated with multiple chromosomes. The downstream utility of reagents generated by the reciprocal probing methods is demonstrated in our studies.


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