RESUMEN
<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>
Asunto(s)
Animales , Femenino , Humanos , Masculino , Ratones , Secuencia de Aminoácidos , Apoptosis , Genética , Secuencia de Bases , Clonación Molecular , ADN Complementario , Química , Genética , Expresión Génica , Proteínas de Choque Térmico , Datos de Secuencia Molecular , Proteínas , Genética , Alineación de Secuencia , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Espermatocitos , Biología Celular , MetabolismoRESUMEN
<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>
Asunto(s)
Niño , Humanos , Masculino , Sustitución de Aminoácidos , Secuencia de Bases , China , Codón sin Sentido , ADN , Química , Genética , Análisis Mutacional de ADN , Iduronato Sulfatasa , Genética , Mucopolisacaridosis II , Genética , Mutación , Mutación Puntual , Polimorfismo Conformacional Retorcido-SimpleRESUMEN
<p><b>OBJECTIVES</b>To evaluate the relationship between microdeletion or mutation on the Y chromosome and Chinese patients with idiopathic azoospermia and severe oligozoospermia and to establish a molecular detection method.</p><p><b>METHODS</b>Microdeletion or mutation detection at the AZFa (sY84 and USP9Y), AZFb, AZFc/DAZ and SRY regions of the Y chromosome. Seventy-three azoospermia and 28 severe oligozoospermia patients were evaluated using PCR and PCR-SSCP techniques.</p><p><b>RESULTS</b>Twelve of 101 patients (12%) with the AZFc/DAZ microdeletion were found, including 8 with azoospermia (11%) and 4 with severe oligozoospermia (14.3%), and 1 patient had a AZFb and AZFc/DAZ double deletion. No deletions in the AZFa or SRY regions were found. No deletions in AZFa, AZFb, AZFc/DAZ or SRY regions were found in 60 normal men who had produced one or more children.</p><p><b>CONCLUSIONS</b>Microdeletion on the Y chromosome, especially at its AZFc/DAZ regions, may be a major cause of azoospermia and severe oligozoospermia leading to male infertility in China. It is recommended that patients have genetic counseling and microdeletion detection on the Y chromosome before intracytoplasmic sperm injection.</p>
Asunto(s)
Humanos , Masculino , Deleción Cromosómica , Oligospermia , Genética , Inyecciones de Esperma Intracitoplasmáticas , Cromosoma YRESUMEN
Two-dimensional polyacrylamide gel electrophoresis (2D PAGE) followed by mass spectrometry (MS) is the most widely used method of protein resolution and identification. But it exist shortcomings. In this paper introduce some novel methods for protein resolution and identification,including the methods by high-liquid chromotography,capillary isoelectric focusing,capillary electrophoresis or 1D and 2D microcapillary chromotagraphy.