Enpeng Zhao

Identification of a novel human angiopoietin-like gene expressed mainly in heart

AbstractThe angiopoietins are an important family of growth factors specific for vascular endothelium. Most of them bind to the TIE2 receptor and are related to regulation of angiogenesis. During large-scale DNA sequencing of the human fetal brain cDNA library, we cloned a novel human angiopoietin-like cDNA and termed it human angiopoietin-like 5 (ANGPTL5). Like other members of the angiopoietin family, ANGPTL5-deduced protein also has an N-terminal cleavable signal peptide, a predicted coiled-coil domain, and a fibrinogen-like domain. The search against the human genome database indicated that ANGPTL5 maps to 11q22. Expression analysis of ANGPTL5 shows that it is mainly expressed in adult human heart.

Identification of a novel human doublecortin-domain-containing gene ( DCDC1 ) expressed mainly in testis

AbstractMutations in the X-linked gene doublecortex (DCX) result in lissencephaly in males or subcortical laminar heterotopia (double cortex) in females. Recently, an evolutionarily conserved doublecortin (DC) domain important for microtubule binding and microtubule polymerization was defined according to detailed sequence analysis of DCX and DCX-like proteins. Subsequently we cloned a novel human cDNA that contained a DC domain during large-scale DNA sequencing of the human fetal brain cDNA library, and termed it doublecortin-domain-containing 1 (DCDC1). According to a search against the human genome database, DCDC1 was mapped to 11p13. Expression analysis showed that DCDC1 was mainly expressed in adult testis. Furthermore, the expression level of DCDC1 in fetal brain was much higher than in adult brain.

A novel human gene whose product shares homology with bovine brain-specific protein p25 is expressed in fetal brain but not in adult brain.

AbstractWe have cloned a novel human gene (C14orf5) from a fetal brain cDNA library that is located on chromosome 14 and consists of 4 exons. It encodes a protein of 170 amino acids that shares homology with human p25 alpha and bovine p25. Reverse transcription-polymerase chain reaction analysis indicated that it is highly expressed in liver and pancreas. Its transcripts could not be detected in adult brain but could be found in fetal brain.

Cloning and Expression of Human GTDC1 Gene (Glycosyltransferase-Like Domain Containing 1) from Human Fetal Library

The glycosyltransferases (GTs) catalyze the synthesis of the carbohydrate portions of glycoproteins, glycolipids, and proteoglycans. Here we report the cloning and characterization of a novel human GTDC1 (glycosyltransferase-like domain containing 1) gene, which locates on human chromosome 2q22. The GTDC1 cDNA is 2954 bp in length, encoding a putative protein of 458 amino acids. At protein level human GTDC1 has 75 and 37% identity with its homologous counterparts in the mouse and fruitfly, respectively. RT-PCR analysis revealed its relatively high expression level in the adult lung, spleen, testis, and peripheral blood leukocyte.

Isolation and identification of a novel cDNA that encodes human yrdC protein

AbstractIn the course of detecting the interaction protein of RBBP10 by yeast two-hybridization, we isolated a novel cDNA that encodes a putative human protein with yrdC domain. It is named human yrdC protein. Because the cDNA contains an open reading fragment (ORF) without a 5′ in- frame stop codon, 5′ RACE and 3′ RACE were proceeded to produce the full-length cDNA. An 1825 bp cDNA was isolated from human placenta, which encodes a putative protein of 279 amino acids. The protein contains a sua5-yciO-yrdC domain. Blast analysis against the human genome database of Genbank revealed that the gene contains five exons, and assigned the gene to human chromosome 1p34.2. A transcript about 2.5 kb is ubiquitously expressed in human tissues. The gene is highly conserved during evolution.

Cloning and identification of a novel human RNPC3 gene that encodes a protein with two RRM domains and is expressed in the cell nucleus.

The RNA recognition motifs (RRM) domain is one of the most common eukaryotic protein folds. Proteins containing RRM domains function in important steps of posttranscriptional regulation of gene expression and are involved in processing and transport of mRNA precursors. Here we describe the cloning and characterization of a novel human RNPC3 gene containing two RNA recognition motifs. The 1870 bp cDNA encodes a protein with 517 amino acids. It also contains two bipartite nuclear targeting sequences, which is important for nuclear targeting for proteins, especially those functioning in the cell nucleus. The GFP location of the RNPC3 gene product shows that this protein is located in the cell nucleus. RT-PCR reveals that it is abundantly expressed in kidney and pancreas.

Cloning and identification of a novel cDNA which encodes a putative protein with a DnaJ domain and a thioredoxin active motif, human macrothioredoxin.

A 3345 bp cDNA was isolated from the fetal brain cDNA library by high throughput cDNA sequencing. The cDNA with an open reading fragment (ORF) of 2241 bp encodes a 747 amino acids putative protein with a DnaJ N-terminus domain and four thioredoxin active sets. So it is named human macrothioredoxin (hMTHr). The nucleotide sequences reported in this paper have been deposited in the Genbank database under accession number AF490904 and AY089971. We used Northern blot to detect a band with a length of about 5 kb, which was ubiquitously expressed in human adult tissues with different intensities. The expression pattern was verified by RT-PCR, revealing that the transcripts were ubiquitously expressed in fetal tissues and human tumor tissues also. The transcripts in fetal tissues were more numerous than in adult tissues. The transcripts were high in adult testis, adult pancreas, fetal thymus, and fetal kidney. We have found three splice isoforms with lengths of 3483, 3345, and 2982 bp in the sequencing analysis of the RT-PCR products. They encode three putative proteins with 793, 747, and 275 amino acids, respectively. The first two putative proteins contain a DnaJ domain and four thioredoxin domains. The third putative protein only contains an N-terminus DnaJ domain and one thioredoxin active set. Blast analysis against the NCBI database revealed that the gene spans a genome sequence of about 75 kb that contains at least 25 exons and is located in human chromosome 2q32.1. The organization of the functional motifs of hMTHr suggests that the protein might be a member of a molecular chaperone family.

A novel cDNA encodes a putative hRALY-like protein, hRALYL

High throughput cDNA sequencing and 5′-rapid amplification of cDNA ends (5′RACE) isolated two cDNAs that shared the same open reading fragment (ORF). Northern blot analysis with the fetal brain mRNA blots detected two transcripts with the length of 3.2 kb and 2.2 kb respectively. The ORF encodes a 291 residues putative protein that shares great homology with hRALY and hnRNPC. So it was named hRALY like protein, hRALYL. Expression pattern was detected by multiple-issue cDNA (MTC) panel based RT-PCR. It revealed that the transcripts of hRALYL were expressed ubiquitously in human tissues with different intensities. The transcript is highest in brain. Blast analysis found the cDNA corresponding to a contig NT_008292, which revealed the gene containing at least 9 exons and located the gene on human chromosome 8q21.13-8q21.2. hRALYL might be a member of hnRNPC subfamily.

cDNA cloning and expression analysis of a novel human F-box domain containing gene†

F-box proteins are a large family of eukaryotic proteins that are characterized by an approximately 40 amino acid motif. Some F-box proteins are critical for the controlled degradation of cellular regulatory proteins. Here we report that a novel member of F-box proteins, FBXO35 gene, was cloned and identified during the large-scale sequencing analysis from a human fetal brain cDNA library. FBXO35 gene shares amino acid similarity with several putative mouse genes not only in F-box domain but also in the rest of the sequence, which indicates that FBXO35 might also contain some other unknown conserved domain. RT-PCR analysis indicated that FBXO35 gene had a ubiquitously low expression pattern in most human adult tissues. According to bioinformatics analysis, the FBXO35 gene was found located in chromosome 3p21.

Cloning and characterization of human synaptotagmin 10 gene.

Synaptotagmin (Syt) is a membrane protein family of secretory vesicles, abundant in neural and some endocrine cells. All the members of this family contain one transmembrane region and two conserved C2 domains. Here we reported a new human Syt 10 gene isolated when screening a human fetal brain cDNA library. This cDNA clone is 3287 bp and contains an open reading frame from 299 to 1870 encoding a putative protein of 523 amino acids. It shares 94.6 and 94.8% homology to rat Syt 10 and mouse Syt 10 at protein level, respectively. RT-PCR result showed that it is expressed only in pancreas, lung and kidney.