RESUMO
<p><b>OBJECTIVE</b>To clone a novel gene which is related to human testis spermatogenesis apoptosis.</p><p><b>METHODS</b>To rapidly attain human novel gene full-length cDNA sequence from a human testis cDNA library,the gene-specific primers and the vector-specific primers were designed for nested polymerase chain reaction. Sequencing was performed and the result was analysed.</p><p><b>RESULTS</b>The present authors discovered the TSARG3 gene(GenBank accession number AF419291) from a human testis cDNA library, using a cDNA fragment (GenBank accession number BE644537) as an electronic probe, which was significantly changed in cryptorchidism and represented a novel gene. Furthermore, a mouse homologue of this gene was identified (GenBank accession number AF419292) by using the same method.</p><p><b>CONCLUSION</b>A novel gene named TSARG3 was cloned. It is considered that the function of the new gene is related to human testis spermatogenesis apoptosis.</p>
Assuntos
Animais , Feminino , Humanos , Masculino , Camundongos , Sequência de Aminoácidos , Apoptose , Genética , Sequência de Bases , Clonagem Molecular , DNA Complementar , Química , Genética , Expressão Gênica , Proteínas de Choque Térmico , Dados de Sequência Molecular , Proteínas , Genética , Alinhamento de Sequência , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Espermatócitos , Biologia Celular , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To investigate and establish the gene diagnosis methods for the frequent mutations of iduronate-2-sulphatase(IDS) gene in mucopolysaccharidosis type II patients.</p><p><b>METHODS</b>polymerase chain- reaction-single strand conformation polymorphism PCR-SSCP) analysis was applied to detect the mutations of exons 3, 8 and 9 which were hot spots in the iduronate-2-sulfatase gene; DNA sequencing was applied to analyze the mutations which had been detected by PCR-SSCP; PCR-restriction fragment length polymorphism (PCR-RFLP) was applied to detect the results of DNA sequencing.</p><p><b>RESULTS</b>Obvious and abnormal bands in exon 9 of the IDS gene were found by applying PCR-SSCP; the mutation(C1672T) of exon 9 was found in the patient through DNA sequencing, which led to amino acid replacement(R468W); the PCR-restriction enzyme digestion showed that only one band(554 bp) appeared in the patient, but there were two bands (257 bp and 297 bp) in his parents, and it verified the results of sequencing analysis.</p><p><b>CONCLUSION</b>PCR-SSCP analysis, DNA sequencing analysis and PCR-restriction enzyme digestion are effective methods for MPS II diagnosis. Combined applications of these methods can verify and complement each other and improve the accuracy of diagnosis.</p>