Isolation of cosmids and fetal brain cDNAs from the proximal long arm of human chromosome 22.
The proximal portion of human chromosome 22q appears to carry genes implicated in the pathogenesis of various developmental disorders, including the cat eye syndrome (CES) and the DiGeorge syndrome (DGS). A cosmid library was prepared from a radiation hybrid selected for its content in chromosome 22 fragments. A large fraction of cosmids containing human DNA were found to derive from the juxtacentromeric region of chromosome 22, as shown by fluorescence in situ hybridization (FISH) performed using individual cosmids or cosmid pools as probes. Finer mapping was obtained for individual cosmids by hybridization to a somatic cell hybrid mapping panel which splits the long arm of the chromosome into 14 bins numbered 1 to 14 from the centromere to the telomere. Of the 10 cosmids mapped, eight belonged to group 1, the other two to group 14, in agreement with FISH data. Rare endonuclease sites and fragments conserved between species were searched in single cosmids, resulting in the selection of seven cosmid fragments which were used to screen a human fetal brain cDNA library. Three cDNAs were identified, encoded from two chromosome 22 genes which appeared to be novel, as determined from partial end sequence and comparison with the database entries. Fine localization of the 30.9 cDNA indicated that the corresponding gene was located in a segment of proximal 22q overlapping with the critical DGS region.
Lamour
V
V
Laboratoire de Biologie des Tumeurs Humaines, CNRS URA 1156, Institut Gustave Roussy, Villejuif, France.
Lévy
N
N
Desmaze
C
C
Baude
V
V
Lécluse
Y
Y
Delattre
O
O
Bernheim
A
A
Thomas
G
G
Aurias
A
A
Lipinski
M
M
D000818:Animals; D001921:Brain / Q000378:metabolism; D002892:Chromosomes, Human, Pair 22; D003360:Cosmids / Q000235:genetics / Q000302:isolation & purification*; D006224:Cricetinae; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D004062:DiGeorge Syndrome / Q000235:genetics; D005333:Fetus / Q000378:metabolism; D015723:Gene Library; D005822:Genetic Vectors; D015894:Genome, Human; D006801:Humans; D006822:Hybrid Cells; D051379:Mice
D004247:DNA