目的为识别参考个体中存在的可疑半同胞,在没有父母遗传信息情况下,解决可疑同胞与参考个体之间的全同胞及半同胞复杂亲缘关系鉴定问题。方法通过检测常染色体并计算参考同胞之间的全同胞指数(Full-sib Index,FSI)及进行IBS评分(Identit...目的为识别参考个体中存在的可疑半同胞,在没有父母遗传信息情况下,解决可疑同胞与参考个体之间的全同胞及半同胞复杂亲缘关系鉴定问题。方法通过检测常染色体并计算参考同胞之间的全同胞指数(Full-sib Index,FSI)及进行IBS评分(Identity by states score),并检测Y染色体以进一步明确参考同胞中的可疑半同胞;根据参考同胞信息构建亲代X染色体及常染色体基因型,并利用已识别的参考半同胞信息推导母亲常染色体基因型。结果确定了参考个体中一名半同胞,最后在没有线粒体技术辅助的情况下,不仅排除了评估对象与参考个体之间的全同胞关系,也明确否定了半同胞关系。结论充分利用参考个体(本案中的全同胞与半同胞)遗传信息,能为复杂亲缘鉴定提供有力的判定依据。展开更多
Both transcription and post-transcriptional processes, such as alternative splicing, play crucial roles in controlling developmental programs in metazoans. Recently emerged RNA-seq method has brought our understanding...Both transcription and post-transcriptional processes, such as alternative splicing, play crucial roles in controlling developmental programs in metazoans. Recently emerged RNA-seq method has brought our understanding of eukaryotic transcriptomes to a new level, because it can resolve both gene expression level and alternative splicing events simultaneously. To gain a better understanding of cellular differentiation in gonads, we analyzed mRNA profiles from Drosophila testes and ovaries using RNA-seq. We identified a set of genes that have sex-specific isoforms in wild-type （WT） gonads, including several transcription factors. We found that differentiation of sperms from undifferentiated germ cells induced a dramatic downregulation of RNA splicing factors. Our data confirmed that RNA splicing events are significantly more frequent in the undifferentiated cell-enriched bag of marbles （barn） mutant testis, but downregulated upon differentiation in WT testis. Consistent with this, we showed that genes required for meiosis and terminal differentiation in WT testis were mainly regulated at the transcriptional level, but not by alternative splicing. Unexpectedly, we observed an increase in expression of all families of chromatin remodeling factors and histone modifying enzymes in the undifferentiated cell-enriched bam testis. More interestingly, chromatin regulators and histone modifying enzymes with opposite enzymatic activities are coenriched in undifferentiated cells in testis, suggesting that these cells may possess dynamic chromatin architecture. Finally, our data revealed many new features of the Drosophila gonadal transcriptomes, and will lead to a more comprehensive understanding of how differential gene expression and splicing regulate gametogenesis in Drosophila. Our data provided a foundation for the systematic study of gene expression and alternative splicing in many interesting areas of germ cell biology in Droso- phila, such as the molecular basis for sexual dimorphism and the regulation of the proliferation vs terminal differentiation programs in germline stem cell lineages. The GEO accession number for the raw and analyzed RNA-seq data is GSE16960.展开更多
To study the expression characteristic of Japanese encephalitis virus (JEV) prME and E proteins and the efficacy of DNA immunization by different recombinant plasmids containing JEV prME (2001 bp) and E (1500 bp) gene...To study the expression characteristic of Japanese encephalitis virus (JEV) prME and E proteins and the efficacy of DNA immunization by different recombinant plasmids containing JEV prME (2001 bp) and E (1500 bp) genes, two recombinants (pJME and pJE) containing JEV prME and E genes fused with FLAG were constructed and then transfected into HepG2 and COS-1 cells by liposome fusion. The expression feature of FLAG-prME (about 72 kDa) and FLAG-E (about 54 kDa) proteins in transfected cells were analyzed by Western blot and two antibody systems (anti-FLAG and anti-E). BALB/c mice were immunized with 100 μg of two kinds of recombinants by intramuscular injection, and JEV JaGAr-01 strains (10 5 PFU/100 μl)were given to BALB/c mice by intraperioneal injection 3 wk after twice DNA immunization by a lethal virus challenge. BALB/c mice were observed for 21 days after challenge. 80% plaque reduction neutralization test was performed to titrate neutralization antibody before and after viral challenge. It was found that the expression of proteins associated with pJME and pJE was determined in transfected cells with anti-FLAG and a new protein of 11 kDa was detected in HepG2 and COS-1 cells transfected with pJME. Only E (53 kDa) protein was identified as transfected with pJME using anti-E. Higher level of neutralization antibodies and the efficacy of protective immunity were induced with pJME immunization, and were similar to those induced by inactivated Japanese encephalitis vaccine, but were better than those induced with pJE. It concludes that the expression level from prM to E proteins of JEV is different in vitro, and the in vitro expression efficiency of pJME was better than that of pJE. FLAG-prME protein expressed by pJME could be cleaved by peptidase from host. The efficacy of DNA immunization is correlated to the expression characterization of related proteins expressed in vitro.展开更多
Atrial fibrillation （AF） remains one of the leading causes of morbidity and mortality in the world which are related to palpita- tions, fainting, congestive heart failure or stroke. The mechanism for atria/fibrillat...Atrial fibrillation （AF） remains one of the leading causes of morbidity and mortality in the world which are related to palpita- tions, fainting, congestive heart failure or stroke. The mechanism for atria/fibrillation has been identified as electrical remod- eling, structure remodeling and intracellular calcium handling remodeling, microRNAs （miRNAs） have recently emerged as one of the important factors in regulating gene expression. So far, thousands of miRNA genes have been found in diverse ani- mals with the function of regulating cell death, cell proliferation, haematopoiesis and even participate in the processing of car- diovascular disease. In this review, we summarize the mechanism of AF and the association of microRNAs network with AF. We provide a potential perspective of miRNAs as the therapeutic target for AF.展开更多
文摘目的为识别参考个体中存在的可疑半同胞,在没有父母遗传信息情况下,解决可疑同胞与参考个体之间的全同胞及半同胞复杂亲缘关系鉴定问题。方法通过检测常染色体并计算参考同胞之间的全同胞指数(Full-sib Index,FSI)及进行IBS评分(Identity by states score),并检测Y染色体以进一步明确参考同胞中的可疑半同胞;根据参考同胞信息构建亲代X染色体及常染色体基因型,并利用已识别的参考半同胞信息推导母亲常染色体基因型。结果确定了参考个体中一名半同胞,最后在没有线粒体技术辅助的情况下,不仅排除了评估对象与参考个体之间的全同胞关系,也明确否定了半同胞关系。结论充分利用参考个体(本案中的全同胞与半同胞)遗传信息,能为复杂亲缘鉴定提供有力的判定依据。
文摘Both transcription and post-transcriptional processes, such as alternative splicing, play crucial roles in controlling developmental programs in metazoans. Recently emerged RNA-seq method has brought our understanding of eukaryotic transcriptomes to a new level, because it can resolve both gene expression level and alternative splicing events simultaneously. To gain a better understanding of cellular differentiation in gonads, we analyzed mRNA profiles from Drosophila testes and ovaries using RNA-seq. We identified a set of genes that have sex-specific isoforms in wild-type （WT） gonads, including several transcription factors. We found that differentiation of sperms from undifferentiated germ cells induced a dramatic downregulation of RNA splicing factors. Our data confirmed that RNA splicing events are significantly more frequent in the undifferentiated cell-enriched bag of marbles （barn） mutant testis, but downregulated upon differentiation in WT testis. Consistent with this, we showed that genes required for meiosis and terminal differentiation in WT testis were mainly regulated at the transcriptional level, but not by alternative splicing. Unexpectedly, we observed an increase in expression of all families of chromatin remodeling factors and histone modifying enzymes in the undifferentiated cell-enriched bam testis. More interestingly, chromatin regulators and histone modifying enzymes with opposite enzymatic activities are coenriched in undifferentiated cells in testis, suggesting that these cells may possess dynamic chromatin architecture. Finally, our data revealed many new features of the Drosophila gonadal transcriptomes, and will lead to a more comprehensive understanding of how differential gene expression and splicing regulate gametogenesis in Drosophila. Our data provided a foundation for the systematic study of gene expression and alternative splicing in many interesting areas of germ cell biology in Droso- phila, such as the molecular basis for sexual dimorphism and the regulation of the proliferation vs terminal differentiation programs in germline stem cell lineages. The GEO accession number for the raw and analyzed RNA-seq data is GSE16960.
基金This research was supported by a grant for project research from high Technology center of Kanazawa Medical University(H2000 2)
文摘To study the expression characteristic of Japanese encephalitis virus (JEV) prME and E proteins and the efficacy of DNA immunization by different recombinant plasmids containing JEV prME (2001 bp) and E (1500 bp) genes, two recombinants (pJME and pJE) containing JEV prME and E genes fused with FLAG were constructed and then transfected into HepG2 and COS-1 cells by liposome fusion. The expression feature of FLAG-prME (about 72 kDa) and FLAG-E (about 54 kDa) proteins in transfected cells were analyzed by Western blot and two antibody systems (anti-FLAG and anti-E). BALB/c mice were immunized with 100 μg of two kinds of recombinants by intramuscular injection, and JEV JaGAr-01 strains (10 5 PFU/100 μl)were given to BALB/c mice by intraperioneal injection 3 wk after twice DNA immunization by a lethal virus challenge. BALB/c mice were observed for 21 days after challenge. 80% plaque reduction neutralization test was performed to titrate neutralization antibody before and after viral challenge. It was found that the expression of proteins associated with pJME and pJE was determined in transfected cells with anti-FLAG and a new protein of 11 kDa was detected in HepG2 and COS-1 cells transfected with pJME. Only E (53 kDa) protein was identified as transfected with pJME using anti-E. Higher level of neutralization antibodies and the efficacy of protective immunity were induced with pJME immunization, and were similar to those induced by inactivated Japanese encephalitis vaccine, but were better than those induced with pJE. It concludes that the expression level from prM to E proteins of JEV is different in vitro, and the in vitro expression efficiency of pJME was better than that of pJE. FLAG-prME protein expressed by pJME could be cleaved by peptidase from host. The efficacy of DNA immunization is correlated to the expression characterization of related proteins expressed in vitro.
基金supported by the National Natural Science Foundation of China (Grant No. 81100122) to Zhang YingChina Postdoctoral Special Science Foundation to Dong DeLiNational Basic Research Program of China (Grant Nos. 2007CB512000 and 2007CB512006) to Yang BaoFeng
文摘Atrial fibrillation （AF） remains one of the leading causes of morbidity and mortality in the world which are related to palpita- tions, fainting, congestive heart failure or stroke. The mechanism for atria/fibrillation has been identified as electrical remod- eling, structure remodeling and intracellular calcium handling remodeling, microRNAs （miRNAs） have recently emerged as one of the important factors in regulating gene expression. So far, thousands of miRNA genes have been found in diverse ani- mals with the function of regulating cell death, cell proliferation, haematopoiesis and even participate in the processing of car- diovascular disease. In this review, we summarize the mechanism of AF and the association of microRNAs network with AF. We provide a potential perspective of miRNAs as the therapeutic target for AF.
