Molecular Cloning and Expression of the Human Homologue of the Murine Gene Encoding Myeloid Leukemia-Inhibitory Factor

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 85; no. 8; pp. 2623 - 2627
• Main Authors: Gough, Nicholas M., Gearing, David P., King, Julie A., Willson, Tracy A., Hilton, Douglas J., Nicola, Nicos A., Metcalf, Donald
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01-04-1988; National Acad Sciences
• Subjects: Amino Acid Sequence; Amino acids; Base Sequence; Biological and medical sciences; Biological Products - genetics; Biotechnology; Cell Differentiation; Cell lines; Cells; Cellular differentiation; Cellular receptors; Cloning, Molecular; Complementary DNA; Cytokines; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation; Genetic engineering; Genetic technics; Growth Inhibitors - genetics; Humans; Interleukin-6; Leukemia Inhibitory Factor; Leukemia, Myeloid, Acute - pathology; Lymphokines; Methods. Procedures. Technologies; Mice; Molecular cloning; Molecular Sequence Data; Monocytes - cytology; Nucleic Acid Hybridization; Proteins; Saccharomyces cerevisiae; Stem cells; Yeasts; Human; Rodentia; Hematopoietic cell; Homology; Primary structure; Cell differentiation; Gene expression; Vertebrata; Mammalia; Gene; Mouse; Analog; Molecular cloning
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.85.8.2623

A human homologue of the recently cloned murine leukemia-inhibitory factor (LIF) gene was isolated from a genomic library by using the murine cDNA as a hybridization probe. The nucleotide sequence of the human gene indicated that human LIF has 78% amino acid sequence identity with murine LIF, with no insertions or deletions, and that the region of the human gene encoding the mature protein has one intervening sequence. After oligonucleotide-mediated mutagenesis, the mature protein-coding region of the LIF gene was introduced into the yeast expression vector YEpsec1. Yeast cells transformed with the resulting recombinant could be induced with galactose to produce high levels of a factor that induced the differentiation of murine M1 leukemic cells in a manner analogous to murine LIF. This factor competed with 125I-labeled native murine LIF for binding to specific cellular receptors on murine cells, compatible with a high degree of structural similarity between the murine and human factors.