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PMID: 6953442 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Construction and characterization of genomic libraries from specific human chromosomes.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 79 ·No. 9 ·1982-05-00 ·Pages 2971-5

Krumlauf R, Jeanpierre M, Young BD

Abstract

Highly purified fractions of human chromosomes 21 and 22 were isolated from a suspension of metaphase chromosomes stained with ethidium bromide by using a fluorescence-activated cell sorter (FACS II). Two recombinant DNA libraries, representing chromosomes 21 and 22, were constructed by complete digestion of DNA from these fractions with EcoRI and insertion into the vector lambda gtWES lambda B. Twenty clones selected at random from the chromosome 22 library hybridized to EcoRI-digested human DNA, and five of these clones hybridized to single bands identical in size to the phage inserts. These five single-copy sequences and a clone coding for an 8S RNA isolated by screening the chromosome 22 library for expressed sequences were characterized in detail. Hybridization of all six clones to a panel of sorted chromosomes and hybrid cell lines confirmed the assignment of the sequences to chromosome 22. The sequences were localized to regions of chromosome 22 by hybridization to translocated chromosomes sorted from a cell line having a balanced translocation t(17;22)(p13;q11) and to hybrid cell lines containing the various portions of another translocation t(X;22)(q13;q112). Five clones reside on the long arm of chromosome 22 between q112 and pter, while one clone and an 18S rRNA gene isolated from the chromosome 22 library reside pter and g112. The construction of chromosome-specific libraries by this method has the advantage of being direct and applicable to nearly all human chromosomes and will be important in molecular analysis of human genetic diseases.

MeSH Terms
Cell Line Chromosome Mapping/methods Chromosomes, Human Chromosomes, Human, 21-22 and Y Cloning, Molecular/methods DNA, Recombinant Flow Cytometry/methods Humans
Chemicals
DNA, Recombinant
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Krumlauf R
Jeanpierre M
Young B D
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28 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.79.9.2971
Published
1982-05-00
Pages
2971-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC346330
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.79.9.2971 PMC Full Text View journal details
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