Molecular cloning of a novel human testis-specific gene TDRG1 / 南方医科大学学报
Journal of Southern Medical University
;
(12): 631-634, 2009.
Artigo
em Chinês
| WPRIM
| ID: wpr-233724
ABSTRACT
<p><b>OBJECTIVE</b>To clone a novel human testis-specific gene TDRG1.</p><p><b>METHODS</b>A new contig of expression sequence tags (ESTs) Hs.180197 was identified from the testis libraries using digital differential display (DDD) to screen the novel human testis-specific gene. To validate the use of bioinformatics approaches in gene discovery, the ESTs Hs.180197, which was predicted to be testis specific, was chosen for further study. Reverse transcriptase-polymerase chain reaction (RT-PCR) was performed on different normal tissues to identify the expression of Hs.180197 in human testis. Using bioinformatics methods and IMAGE cloning of this contig, the full-length cDNA sequence of the noval human gene was cloned.</p><p><b>RESULTS</b>This novel gene was 1197 bp in length, located in chromosome 6p21.1-p21.2. The sequence of the open reading frame was 504-806 bp, as confirmed by RT-PCR and sequencing in human testis. The cDNA encodes a novel protein of 100 amino acids with a theoretical molecular weight of 10 000 and isoelectric point of 6.81. The sequence shares no significant homology with any known protein in the databases. Semi-quantitative RT-PCR analysis of multiple tissues further showed that the novel gene was expressed specifically in adult human testis. Considering a possible relation of this novel gene with the function of human testis, we named this new gene TDRG1 (testis development related gene 1, GenBank accession number DQ168992).</p><p><b>CONCLUSION</b>DDD combined with laboratory validation is an efficient method for identifying new human functional genes.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Especificidade de Órgãos
/
Testículo
/
Dados de Sequência Molecular
/
Proteínas
/
Regulação da Expressão Gênica
/
Alinhamento de Sequência
/
Clonagem Molecular
/
DNA Complementar
/
Proteínas de Homeodomínio
/
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Limite:
Adulto
/
Humanos
/
Masculino
Idioma:
Chinês
Revista:
Journal of Southern Medical University
Ano de publicação:
2009
Tipo de documento:
Artigo
Similares
MEDLINE
...
LILACS
LIS