46,XX男性综合征是一类罕见的性发育障碍性疾病,通常染色体为46,XX且Y染色体性别决定区(sex-determining region Y gene,SRY)阳性,有正常的外生殖器和男性表型,多因成年后不育就诊。而SRY阴性的46,XX男性综合征患者多伴发外生殖器发育畸...46,XX男性综合征是一类罕见的性发育障碍性疾病,通常染色体为46,XX且Y染色体性别决定区(sex-determining region Y gene,SRY)阳性,有正常的外生殖器和男性表型,多因成年后不育就诊。而SRY阴性的46,XX男性综合征患者多伴发外生殖器发育畸形,多因年幼时性发育异常就诊。报告1例SRY阴性的46,XX男性综合征患者,该患者外生殖器及第二性征发育均为正常男性型,检查发现患者核型为46,XX、SRY阴性、AZF区域缺失、高促性腺激素性性腺功能减退症且无精子症,其生育建议行供精人工授精。展开更多
Animal genetic resources are playing a vital role in livestock production and are essential to food security. The present study aims to contribute to a better understanding genetic local sheep breeds and to elucidate ...Animal genetic resources are playing a vital role in livestock production and are essential to food security. The present study aims to contribute to a better understanding genetic local sheep breeds and to elucidate the phylogenetic relationships through the evolution of the SRY gene in four different lineages of Ladoum sheep raised in Senegal. After a brief analysis of genetic diversity, the phylogenetic relationships and molecular dating were inferred through haplotype networks and four phylogenetic reconstruction methods. The different haplotype networks are constructed with NETWORK ver. 5.0.0.0 using the Median-Joining method. Phylogenetic trees were reconstructed using neighbor-joining, maximum parsimony, maximum likelihood and Bayesian inference. The robustness of the nodes in phylogenetic trees of the three first methods was assessed by 1000 bootstraps. For Bayesian inference, the posterior probability distribution of the trees was estimated by 4 MCMC chains. 5,000,000 generations were performed for each of the chains by sampling the different parameters every 1000 generations. Results show a low polymorphism. Haplotypic diversity is much higher than the average nucleotide divergence between all pairs of haplotypes. The majority and central haplotype indicates a close relationship between “Batling” and “Tyson” individuals. “Birahim” lineage is very distinct from the rest. Phylogenetic trees confirm two genetically separate clades between “Birahim” and the other lineages. The period of divergence between “Birahim” lineage versus the common ancestor of the other three lineages was 2504 years ago. The polyphyly revealed in “Birahim” lindicates that this lineage does not contain the common ancestor of all individuals who compose it. It could therefore be derived from two or more sheep breeds with a common ancestor, Ovis aries. The monophyletic clade appears to be a group including a common ancestor and all of its genetic descendants. This group, bringing together the other three lineages, is in the process of being structured into sub-lineages. This study is the first to show that there are only two genetic lines within ladoum sheep in Senegal.展开更多
Background: In disorders of sexual differentiation, sexual development may not conform to the chromosomal structure, thus forming different types of abnormalities. Among these abnormalities is syndrome 46, XX DSD wher...Background: In disorders of sexual differentiation, sexual development may not conform to the chromosomal structure, thus forming different types of abnormalities. Among these abnormalities is syndrome 46, XX DSD where most patients are female phenotype with clitoral hypertrophy that can go to complete masculinization especially in the presence of the SRY gene. Objective: The goal of this work is to demonstrate a relationship between the genotype and the phenotype in five patients karyotype 46, XX with the presence of the SRY gene. Methodology: The study involves five patients referred to the laboratory under suspicion of sexual development anomalies. The diagnosis took place through hormonal and echography examinations, a classic cytogenetic study (Barr chromatin and karyotype) and an amplification of the SRY gene located on the Y chromosome. The resulting PCR products were sent for sequencing. Results: Based on the results of clinical and paraclinical tests carried out it was found clitoral hypertrophy, the presence of clitoris penis for some, presence of normal penis for others. In addition, echography revealed a lack of female internal genitalia (P2, P3), and a presence of testicles (P3, P4, P5). Genetic analysis (chromosomal and molecular) showed a karyotype 46, XX SRY (+) for all patients. New mutations were found c.246 T > A, p.82 Asn82Lys and c.171 G > C, p.57 Gln57His. Conclusion: In our study, we were able to correlate each DSD with karyotype 46, XX to a pathology such as 46, XX DSD testicular, 46, XX DSD with clitoral hypertrophy and ovotestis 46, XX. The next step will undoubtedly be the integration of other molecular techniques (genotyping, FISH, CGH or even the CGH array) to further genetic exploration.展开更多
利用PCR技术从雄性中国荷斯坦牛的基因组DNA中克隆了Y染色体性别决定基因(Sex-deter-m in ing R eg ion on the Y Chrom osom e,SRY)的编码区全长序列,构建了表达载体pET-28a/SRY,并将其在大肠杆菌(E.coli)中进行了诱导表达,对表达产物...利用PCR技术从雄性中国荷斯坦牛的基因组DNA中克隆了Y染色体性别决定基因(Sex-deter-m in ing R eg ion on the Y Chrom osom e,SRY)的编码区全长序列,构建了表达载体pET-28a/SRY,并将其在大肠杆菌(E.coli)中进行了诱导表达,对表达产物进行了检测。结果表明,SRY基因编码区长687 bp,编码229个氨基酸;表达载体pET-28a/SRY构建成功;表达产物中含有相对分子质量为33 ku的SRY蛋白。展开更多
以流式细胞仪分离小麂(Muntiacus reevesi)Y染色体和黑麂(Muntiacus crinifrons)Y1,Y2,X+4和1号染色体,利用DOP-PCR技术富集了分离的各单条染色体。然后,将小麂的Y染色体的DOP-PCR产物经Cy3标记后直接作为涂染探针, 应用染色体涂染技术...以流式细胞仪分离小麂(Muntiacus reevesi)Y染色体和黑麂(Muntiacus crinifrons)Y1,Y2,X+4和1号染色体,利用DOP-PCR技术富集了分离的各单条染色体。然后,将小麂的Y染色体的DOP-PCR产物经Cy3标记后直接作为涂染探针, 应用染色体涂染技术与雌雄黑麂的核型标本进行杂交,确认了黑麂真正的Y染色体为Y2染色体。再以黑麂的Y1,Y2,X+4和1号染色体的DOP-PCR产物为模板,用人的特异性的 SRY（sex determining region of the Y chromosome ） 基因引物对其进行扩增,结果表明黑麂只有Y2染色体出现了SRY扩增片段。然后扩增产物克隆和测序,比较它与人的同源性,初步把黑麂的Sry基因定位在Y2染色体上。最后提取雄性黑麂的基因组DNA,并用同一对引物对其进行扩增,亦得到Sry基因的片段,对此扩增片段进行克隆,测序,结果表明其与Y2染色体得到的Sry基因片段完全一样，与人SRY基因的同源性均为83%。展开更多
We amplified the 197 bp HMG box sequence by using Polymerase Chain Reaction (PCR) with primers according to the known SRY sequence from male raccoon genomic DNA, and then cloned and sequenced. The nucleic acid and ami...We amplified the 197 bp HMG box sequence by using Polymerase Chain Reaction (PCR) with primers according to the known SRY sequence from male raccoon genomic DNA, and then cloned and sequenced. The nucleic acid and amino acid sequence comparison between raccoon and other mammals revealed high conservation of the SRY HMG box in mammals (about 80%), which implied that the DNA binding activity was crucial to mediate the action of SRY in sex determination. The variation of raccoon HMG box sequence mainly occurred on purine by replacement and missense mutation, which implicated that only the HMG box protein with advanced structure had efficient activity. The homology and difference between mammals HMG box was accorded to the evolution systematic tree.展开更多
目的:探讨性反转综合征(sex reversal syndrome,SRS)的发病原因。方法:应用聚合酶链式反应(PCR)及PCR产物直接测序方法对22例性发育异常患者的人类性别决定区域(sex-determining region of Y chromosome,SRY)基因进行特异性扩增和序列...目的:探讨性反转综合征(sex reversal syndrome,SRS)的发病原因。方法:应用聚合酶链式反应(PCR)及PCR产物直接测序方法对22例性发育异常患者的人类性别决定区域(sex-determining region of Y chromosome,SRY)基因进行特异性扩增和序列检测分析。结果:在22例患者中有7例患者SRY基因检测结果与染色体性别不一致,其中3例为46,XX女性性反转患者的SRY基因为阳性(其中1例46,XX,Gp+),3例为46,XY男性性反转患者的SRY缺失,1例为45,X女性患者的SRY基因阳性;其他15例患者SRY基因与染色体性别一致,但都伴有不同程度的性发育异常,即存在Y染色体患者,SRY为阳性;无Y染色体的患者,SRY为阴性。通过对12例SRY阳性患者进行测序分析,均未发现碱基突变。结论:SRY基因的缺失或易位是导致SRS的最主要原因,同时也表明人类的性别决定和分化还有其他相关基因参与。展开更多
文摘46,XX男性综合征是一类罕见的性发育障碍性疾病,通常染色体为46,XX且Y染色体性别决定区(sex-determining region Y gene,SRY)阳性,有正常的外生殖器和男性表型,多因成年后不育就诊。而SRY阴性的46,XX男性综合征患者多伴发外生殖器发育畸形,多因年幼时性发育异常就诊。报告1例SRY阴性的46,XX男性综合征患者,该患者外生殖器及第二性征发育均为正常男性型,检查发现患者核型为46,XX、SRY阴性、AZF区域缺失、高促性腺激素性性腺功能减退症且无精子症,其生育建议行供精人工授精。
文摘Animal genetic resources are playing a vital role in livestock production and are essential to food security. The present study aims to contribute to a better understanding genetic local sheep breeds and to elucidate the phylogenetic relationships through the evolution of the SRY gene in four different lineages of Ladoum sheep raised in Senegal. After a brief analysis of genetic diversity, the phylogenetic relationships and molecular dating were inferred through haplotype networks and four phylogenetic reconstruction methods. The different haplotype networks are constructed with NETWORK ver. 5.0.0.0 using the Median-Joining method. Phylogenetic trees were reconstructed using neighbor-joining, maximum parsimony, maximum likelihood and Bayesian inference. The robustness of the nodes in phylogenetic trees of the three first methods was assessed by 1000 bootstraps. For Bayesian inference, the posterior probability distribution of the trees was estimated by 4 MCMC chains. 5,000,000 generations were performed for each of the chains by sampling the different parameters every 1000 generations. Results show a low polymorphism. Haplotypic diversity is much higher than the average nucleotide divergence between all pairs of haplotypes. The majority and central haplotype indicates a close relationship between “Batling” and “Tyson” individuals. “Birahim” lineage is very distinct from the rest. Phylogenetic trees confirm two genetically separate clades between “Birahim” and the other lineages. The period of divergence between “Birahim” lineage versus the common ancestor of the other three lineages was 2504 years ago. The polyphyly revealed in “Birahim” lindicates that this lineage does not contain the common ancestor of all individuals who compose it. It could therefore be derived from two or more sheep breeds with a common ancestor, Ovis aries. The monophyletic clade appears to be a group including a common ancestor and all of its genetic descendants. This group, bringing together the other three lineages, is in the process of being structured into sub-lineages. This study is the first to show that there are only two genetic lines within ladoum sheep in Senegal.
文摘Background: In disorders of sexual differentiation, sexual development may not conform to the chromosomal structure, thus forming different types of abnormalities. Among these abnormalities is syndrome 46, XX DSD where most patients are female phenotype with clitoral hypertrophy that can go to complete masculinization especially in the presence of the SRY gene. Objective: The goal of this work is to demonstrate a relationship between the genotype and the phenotype in five patients karyotype 46, XX with the presence of the SRY gene. Methodology: The study involves five patients referred to the laboratory under suspicion of sexual development anomalies. The diagnosis took place through hormonal and echography examinations, a classic cytogenetic study (Barr chromatin and karyotype) and an amplification of the SRY gene located on the Y chromosome. The resulting PCR products were sent for sequencing. Results: Based on the results of clinical and paraclinical tests carried out it was found clitoral hypertrophy, the presence of clitoris penis for some, presence of normal penis for others. In addition, echography revealed a lack of female internal genitalia (P2, P3), and a presence of testicles (P3, P4, P5). Genetic analysis (chromosomal and molecular) showed a karyotype 46, XX SRY (+) for all patients. New mutations were found c.246 T > A, p.82 Asn82Lys and c.171 G > C, p.57 Gln57His. Conclusion: In our study, we were able to correlate each DSD with karyotype 46, XX to a pathology such as 46, XX DSD testicular, 46, XX DSD with clitoral hypertrophy and ovotestis 46, XX. The next step will undoubtedly be the integration of other molecular techniques (genotyping, FISH, CGH or even the CGH array) to further genetic exploration.
文摘利用PCR技术从雄性中国荷斯坦牛的基因组DNA中克隆了Y染色体性别决定基因(Sex-deter-m in ing R eg ion on the Y Chrom osom e,SRY)的编码区全长序列,构建了表达载体pET-28a/SRY,并将其在大肠杆菌(E.coli)中进行了诱导表达,对表达产物进行了检测。结果表明,SRY基因编码区长687 bp,编码229个氨基酸;表达载体pET-28a/SRY构建成功;表达产物中含有相对分子质量为33 ku的SRY蛋白。
文摘以流式细胞仪分离小麂(Muntiacus reevesi)Y染色体和黑麂(Muntiacus crinifrons)Y1,Y2,X+4和1号染色体,利用DOP-PCR技术富集了分离的各单条染色体。然后,将小麂的Y染色体的DOP-PCR产物经Cy3标记后直接作为涂染探针, 应用染色体涂染技术与雌雄黑麂的核型标本进行杂交,确认了黑麂真正的Y染色体为Y2染色体。再以黑麂的Y1,Y2,X+4和1号染色体的DOP-PCR产物为模板,用人的特异性的 SRY（sex determining region of the Y chromosome ） 基因引物对其进行扩增,结果表明黑麂只有Y2染色体出现了SRY扩增片段。然后扩增产物克隆和测序,比较它与人的同源性,初步把黑麂的Sry基因定位在Y2染色体上。最后提取雄性黑麂的基因组DNA,并用同一对引物对其进行扩增,亦得到Sry基因的片段,对此扩增片段进行克隆,测序,结果表明其与Y2染色体得到的Sry基因片段完全一样，与人SRY基因的同源性均为83%。
文摘We amplified the 197 bp HMG box sequence by using Polymerase Chain Reaction (PCR) with primers according to the known SRY sequence from male raccoon genomic DNA, and then cloned and sequenced. The nucleic acid and amino acid sequence comparison between raccoon and other mammals revealed high conservation of the SRY HMG box in mammals (about 80%), which implied that the DNA binding activity was crucial to mediate the action of SRY in sex determination. The variation of raccoon HMG box sequence mainly occurred on purine by replacement and missense mutation, which implicated that only the HMG box protein with advanced structure had efficient activity. The homology and difference between mammals HMG box was accorded to the evolution systematic tree.
文摘目的:探讨性反转综合征(sex reversal syndrome,SRS)的发病原因。方法:应用聚合酶链式反应(PCR)及PCR产物直接测序方法对22例性发育异常患者的人类性别决定区域(sex-determining region of Y chromosome,SRY)基因进行特异性扩增和序列检测分析。结果:在22例患者中有7例患者SRY基因检测结果与染色体性别不一致,其中3例为46,XX女性性反转患者的SRY基因为阳性(其中1例46,XX,Gp+),3例为46,XY男性性反转患者的SRY缺失,1例为45,X女性患者的SRY基因阳性;其他15例患者SRY基因与染色体性别一致,但都伴有不同程度的性发育异常,即存在Y染色体患者,SRY为阳性;无Y染色体的患者,SRY为阴性。通过对12例SRY阳性患者进行测序分析,均未发现碱基突变。结论:SRY基因的缺失或易位是导致SRS的最主要原因,同时也表明人类的性别决定和分化还有其他相关基因参与。
