title: Gene transfer, expression, and molecular cloning of the human transferrin receptor gene.
abstract: We describe the molecular cloning of the human transferrin receptor gene by a gene transfer approach. Mouse Ltk- cells were cotransformed with the herpes simplex thymidine kinase gene and total human DNA. Transformants expressing human transferrin receptor were isolated by selection on hypoxanthine/aminopterin/thymidine (HAT) medium and fluorescence-activated cell sorting of HAT-resistant cells. Thirty-four kilobases of human DNA was isolated by screening a genomic library constructed from the DNA of a secondary transformant. Gene transfer of the cloned DNA established that 31 kb of DNA was sufficient to encode the receptor. A probe from the 5' end of the gene was used to isolate a cDNA clone with an insert of 4.9 kb. Hybridization of the cDNA to the cloned genomic DNA revealed a minimum of 12 exons. They extend over the entire 31 kb of expressing DNA and over 2 kb of adjacent 3' untranslated sequences that are not required for receptor expression in L cells.
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004251:DNA Transposable Elements; D005796:Genes; D006801:Humans; D007739:L Cells / Q000378:metabolism; D007945:Leukemia, Lymphoid; D051379:Mice; D009693:Nucleic Acid Hybridization; D014176:Protein Biosynthesis; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000378:metabolism; D011990:Receptors, Transferrin; D013937:Thymidine Kinase / Q000235:genetics; D014168:Transferrin / Q000378:metabolism
chemical_list: D004251:DNA Transposable Elements; D011956:Receptors, Cell Surface; D011990:Receptors, Transferrin; D014168:Transferrin; D013937:Thymidine Kinase; D004262:DNA Restriction Enzymes