Isolation and characterization of a differentiation-dependent gene in the human colonic cell line HT29-18

Archives of Biochemistry and Biophysics
M M OlivaV W Yang

Abstract

The human colonic epithelial cell line HT29, and its clonal derivatives HT29-18 and HT-29-18-C1, differentiate in vitro. Differential screening of a subtraction cDNA library enriched for sequences unique to HT29-18-C1, a highly differentiated subclone of HT29-18, resulted in the isolation of a differentiation-dependent cDNA clone, A4. A full-length clone encoding A4 was obtained and sequenced to its entirety. It is 945 bp in length and contains an open reading frame (ORF) of 456 bp. The amino acid sequence deduced from the ORF reveals a polypeptide of 152 amino acids with a predicted molecular mass of 17,000 Da, a size confirmed by coupled in vitro transcription and translation directed by the full-length A4 cDNA. This polypeptide contains four potential membrane-spanning domains and consensus sequences for N-linked glycosylation as well as phosphorylation sites for protein kinase C and casein kinase II. Comparison of A4 to published DNA and protein sequences revealed no significant homology. Genomic Southern blot analysis suggests that the gene is present in a single copy within the human genome and is conserved in the rat. Northern blot analysis of RNA obtained from various rat tissues shows that the expression of the A4 gene...Continue Reading

Citations

Sep 21, 2005·Genes & Genetic Systems·Takashi Kitano, Naruya Saitou
Oct 12, 2004·Biochemical and Biophysical Research Communications·Sang Min LeeJiyoung Kim
Jul 27, 2001·Mechanisms of Development·S MildeC Kirchner
Aug 30, 2002·Blood·Jaime O ClaudioA Keith Stewart
Jun 26, 2012·Analytical and Bioanalytical Chemistry·Alessandra LongoClaudio Luparello
May 10, 2000·Journal of Biomolecular Screening·M SchusterG Werner
Mar 1, 1997·The American Journal of Physiology·G E BreitwieserV W Yang
Nov 30, 2019·Journal of Cellular and Molecular Medicine·Zichao FengXingang Li

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