Isolation of a series of HLA class I clones from a human chromosome 6 genomic library.
Human metaphase chromosomes were fractionated by a fluorescent-activated cell sorter (FACS II) and the chromosome 6 fraction was sorted. A genomic library was constructed in lambda gtWES cloning arms using a partial EcoRI digest of the chromosomal DNA. We estimate that at least 60% of our library represents chromosome 6 material, and as 1.2 x 10(5) recombinants were obtained, this indicates that the majority of clonable chromosome 6 sequences are represented. The library was screened with a mouse H-2 class I clone and 18 HLA class I recombinants were isolated from 4 x 10(4) plaques.
D002458:Cell Fractionation / Q000379:methods; D002906:Chromosomes, Human, 6-12 and X; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D006680:HLA Antigens / Q000235:genetics*; D006801:Humans
D004274:DNA, Recombinant; D006680:HLA Antigens