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Construction of a DNA Library Enriched with Mouse 4 x Chromosome of T(X;4)37H Translocation
Author(s) -
Takahashi Ichiro,
Ueda Takayuki,
Kameoka Yosuke,
Abe Kenji,
Takagi Nobuo,
Hashimoto Katsuyuki
Publication year - 1990
Publication title -
development, growth and differentiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 66
eISSN - 1440-169X
pISSN - 0012-1592
DOI - 10.1111/j.1440-169x.1990.00321.x
Subject(s) - microbiology and biotechnology , chromosome 22 , biology , chromosomal translocation , chromosome , karyotype , propidium iodide , chromosome 21 , dna , chromosome 16 , chromosome 4 , restriction enzyme , genetics , gene , apoptosis , programmed cell death
Normal mouse chromosomes are routinely separated into only 5 peaks by the current flow cytometry. Since this limited resolution hindered isolation of the normal mouse X chromosome with an appropriate purity, we attempted to sort the mouse 4 x chromosome, a larger translocation chromosome of T(X;4)37H, consisting of nearly the entire chromosome 4 and chromosome X by flow cytometry. To obtain a large number of cells having 4 x chromosome for flow sorting, we isolated a somatic hybrid cell line MHH‐1 formed between S194 myelome cell line and normal splenocytes from a male mouse carrying T(X;4)37H. Flow karyotyping of propidium iodide‐stained chromosomes from MHH‐1 cell line revealed an additional peak containing 4 x chromosomes at about 80%. DNA purified from sorted 4 x chromosomes was cloned into phage lambda gtWES after complete digestion with EcoRl restriction endonuclease. Thus a 4 x chromosome‐enriched library of about 4.4 × 10 4 recombinant phages was made and 13 single copy DNA clones specific to the X chromosome were isolated from the library so far.