Molecular cloning and characterization analysis of HPESCRG1, a novel gene expressed specifically in human embryonic stem cell / 中华医学遗传学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To clone a novel gene expressed specifically in human embryonic stem cell and to analyze its characteristics.</p><p><b>METHODS</b>Based on an expression sequence tags(EST) CF948547 which expressed specifically in human embryonic stem cell, the full-length cDNA sequence of a novel gene was cloned by using bioinformatic and molecular biological technique. Its expression profile was analyzed by reverse transcription-polymerase chain reaction(RT-PCR), and subcellular location was determined by enhanced green fluorescent protein (EGFP) eukaryotic expression system.</p><p><b>RESULTS</b>A novel gene HPESCRG1(homo sapiens pluripotent embryonic stem cell-related gene) was cloned successfully. Its GenBank accession number was AY283672. Its cDNA length was 1395 bp. It comprised 9 exons and 8 introns, and its opening reading frame was 250-1146 bp. Its chromosomal mapping was located in 3q13.13, and the putative protein contained 297 amino acids. The theoretical molecular weight of the putative protein was 33 784 and the isoelectric point was 9.35. The protein primary structure of this gene contained a SAP motif and it was subcellularly located in nuclei. Expression analysis showed that this gene was expressed specifically in human ES cells, but not expressed in the adult human tissues, the multiple tissues of embryo aborted in over 5 months' pregnancy, the differentiated cells of HESC-1, and the human mesenchymal stem cells (hMSCs) and human embryonic fibrocytes (hEFCs).</p><p><b>CONCLUSION</b>HPESCRG1 was found to be a novel gene expressed specifically in human ES cell, which might be related to self-renewal of human ES cell and maintaining its undifferentiated state.</p>