Assignment of tear lipocalin gene to human chromosome 9q34–9qter

Abstract

We assigned the gene for tear lipocalin to the long arm of human chromosome 9. Polyadenylated RNA was extracted from lacrimal gland. The coding region for tear lipocalin was amplified, sequenced and used to probe a panel of somatic cell hybrid DNA by Southern blot analysis. Regional mapping was accomplished by probing a panel of subfragments of the indicated chromosome. Restriction of genomic DNA with EcoRI failed to reveal any bands corresponding to the human tear lipocalin gene in mouse-human hybrids all of which lack chromosome 9. Southern blot analysis of human-hamster hybrids demonstrated a human 5.6 kb TaqI restriction fragment that segregated to the q34-qter region of chromosome 9 and assigned the gene for tear lipocalin to this region. Structurally homologous proteins of the lipocalin family, human placental protein 14, human alpha 1 microglobulin, and human brain prostaglandin synthase, have been mapped to this region. We suggest that the gene for tear lipocalin is part of an important lipocalin superfamily gene cluster on chromosome 9 within band q34.

Assignment of LCN1 to human chromosome 9 is confirmed

“…Using a panel of hybrid cell lines that had been generated and charac terized by Nguyen et al (1986), we mapped the human LCN 1 gene to chromosome 9, supporting the previous conclusion of that the probe, representing the gene in question, maps to chromosome 9. These results confirm the localization, via somatic cell hybrid analysis, of the same tear lipocalin gene by Glasgow et al (1993), although these workers did not use the gene symbol LCN 1.…”

Site-directed circular dichroism of proteins: 1Lb bands of Trp resolve position-specific features in tear lipocalin

“…The TL cDNA in PCR II (Invitrogen, San Diego, CA), previously synthesized [17], was used as a template to clone the TL gene spanning bases 115-592 of the previously published sequence [18] into pET 20b (Novagen). Flanking restriction sites for NdeI and BamHI were added to produce the native protein sequence as found in tears but with an initiating methionine [19].…”

Molten globule state of tear lipocalin: ANS binding restores tertiary interactions

“…The TL cDNA in PCR II previously synthesized [25], was used as a template to clone the TL gene spanning bases 115-592 of the previously published sequence [20] into pET 20b with flanking restriction sites for NdeI and BamHI as previously described [26]. For single Cys mutants of TL, C101L was chosen as the template for additional mutant cDNAs because it showed similar structural features and binding characteristics for spin labeled lipids [27].…”