[ Objective] The aim of this study was to investigate the construction and identification of siRNA expression vector targeting nucleocapsid protein N gone of PRRSV. [Method] Three siRNA oligonucleotides targeting nucl...[ Objective] The aim of this study was to investigate the construction and identification of siRNA expression vector targeting nucleocapsid protein N gone of PRRSV. [Method] Three siRNA oligonucleotides targeting nucleocapsid protein N gone sequence of PRRSV were designed or synthesized, and then inserted into CMV promoter downstream to clone into pSilencer 4,1 -CMV eukaryotic expression vector. The recombinant expression vector was identified by enzyme digestion and DNA sequencing. [ Result] The results showed that the siRNA interference recombinant plasmid vector pSilencer-N targeting nucleocapsid protein gone expression had been successfully constructed. [ Conclusion] This study lays a foundation for studies on the controlling PRRSV by RNA interference technique .展开更多
DNA double-strand breaks （DSBs） are introduced in cells by ionizing radiation and reactive oxygen species. In addition, they are commonly generated during V（D）J recombination, an essential aspect of the developing...DNA double-strand breaks （DSBs） are introduced in cells by ionizing radiation and reactive oxygen species. In addition, they are commonly generated during V（D）J recombination, an essential aspect of the developing immune system. Failure to effectively repair these DSBs can result in chromosome breakage, cell death, onset of cancer, and defects in the immune system of higher vertebrates. Fortunately, all mammalian cells possess two enzymatic pathways that mediate the repair of DSBs： homologous recombination and non-homologous end-joining （NHEJ）. The NHEJ process utilizes enzymes that capture both ends of the broken DNA molecule, bring them together in a synaptic DNA-protein complex, and finally repair the DNA break. In this review, all the known enzymes that play a role in the NHEJ process are discussed and a working model for the co-operation of these enzymes during DSB repair is presented.展开更多
This paper reports a new approach to detect ribozyme cleavage product based on the molecular-beacon-ligation system. The molecular beacon, designed in such a way that one-half of its loop is complementary to ribozyme ...This paper reports a new approach to detect ribozyme cleavage product based on the molecular-beacon-ligation system. The molecular beacon, designed in such a way that one-half of its loop is complementary to ribozyme cleavage product, is used to monitor ligation process of RNA/DNA com-plex in a homogeneous solution and to convert directly cleavage product information into fluorescence signal. The method need not label ribozyme and ribozyme substrate, which is fast, simple and ultra-sensitive for detection of cleavage product. Detection limit of the assay is 0.05 nmol/L. The cleavage product of hammerhead ribozyme against hepatitis C virus RNA (HCV-RNA) was detected perfectly based on this assay. Owing to its ultrasensitivity, excellent specificity, convenience and fidelity, this method might hold out great promise in ribozyme reaction and ribozyme gene therapy.展开更多
Retinoic acid level in the retina/choroid is altered in induced myopia models.All-trans-retinol dehydrogenase(RDH8) is an important enzyme of retinoic acid metabolism.This study aimed to investigate the association of...Retinoic acid level in the retina/choroid is altered in induced myopia models.All-trans-retinol dehydrogenase(RDH8) is an important enzyme of retinoic acid metabolism.This study aimed to investigate the association of the RDH8 gene with high myopia.Three single nucleotide polymorphisms(SNPs) [RDH851(rs2233789) ,RDH8E5a(rs1644731) ,and RDH855b(rs3760753) ]were selected,based on the linkage disequilibrium pattern of RDH8 from a previous study,and genotyped for 160 Han Chinese nuclear families with highly myopic(-10 diopters or worse) offspring as well as in an independent group with 166 highly myopic cases(-10 diopters or worse) and 211 controls. Family-based association analysis was performed using the family-based association test(FBAT) package,and genotype relative risk(GRR) was calculated using the GenAssoc program.Population-based association analysis was performed using Chi-square test.These SNPs were in linkage equilibrium with each other.SNPs RDH851(rs2233789) and RDH8E5a(rs1644731) both did not show association with high myopia.SNP RDH855b(rs3760753) demonstrated significant association(P=0.0269) with a GRR of 0.543(95%confidence interval=0.304-0.968,P=0.038) .The association became statistically insignificant,however,after multiple comparison correction.Haplotype analysis did not show a significant association either.Population-based association analysis also showed no significant association(P>0.05) .Our family-and population-based data both suggest that the RDH8 gene is unlikely to be associated with high myopia in Chinese.展开更多
基金Supported by Based on Cuttingedge technology and research Project of Henan Province(072300430060)The focus of Scientific andTechnological Project of Henan Province(072102130023)Colleges and Universities of Henan Province in Support of TechnologicalInnovation Plan~~
文摘[ Objective] The aim of this study was to investigate the construction and identification of siRNA expression vector targeting nucleocapsid protein N gone of PRRSV. [Method] Three siRNA oligonucleotides targeting nucleocapsid protein N gone sequence of PRRSV were designed or synthesized, and then inserted into CMV promoter downstream to clone into pSilencer 4,1 -CMV eukaryotic expression vector. The recombinant expression vector was identified by enzyme digestion and DNA sequencing. [ Result] The results showed that the siRNA interference recombinant plasmid vector pSilencer-N targeting nucleocapsid protein gone expression had been successfully constructed. [ Conclusion] This study lays a foundation for studies on the controlling PRRSV by RNA interference technique .
文摘DNA double-strand breaks （DSBs） are introduced in cells by ionizing radiation and reactive oxygen species. In addition, they are commonly generated during V（D）J recombination, an essential aspect of the developing immune system. Failure to effectively repair these DSBs can result in chromosome breakage, cell death, onset of cancer, and defects in the immune system of higher vertebrates. Fortunately, all mammalian cells possess two enzymatic pathways that mediate the repair of DSBs： homologous recombination and non-homologous end-joining （NHEJ）. The NHEJ process utilizes enzymes that capture both ends of the broken DNA molecule, bring them together in a synaptic DNA-protein complex, and finally repair the DNA break. In this review, all the known enzymes that play a role in the NHEJ process are discussed and a working model for the co-operation of these enzymes during DSB repair is presented.
基金Supported by the National Key Basic Research Program (Grant No. 2002CB513100)the National Natural Science Foundation of China (Grant Nos. 20505007 and 20475015)the Key Project of Hunan Province Technology Plan of China (Grant Nos. 02JEY2004 and 0399Y1006)
文摘This paper reports a new approach to detect ribozyme cleavage product based on the molecular-beacon-ligation system. The molecular beacon, designed in such a way that one-half of its loop is complementary to ribozyme cleavage product, is used to monitor ligation process of RNA/DNA com-plex in a homogeneous solution and to convert directly cleavage product information into fluorescence signal. The method need not label ribozyme and ribozyme substrate, which is fast, simple and ultra-sensitive for detection of cleavage product. Detection limit of the assay is 0.05 nmol/L. The cleavage product of hammerhead ribozyme against hepatitis C virus RNA (HCV-RNA) was detected perfectly based on this assay. Owing to its ultrasensitivity, excellent specificity, convenience and fidelity, this method might hold out great promise in ribozyme reaction and ribozyme gene therapy.
基金Project supported by the National Natural Science Foundation of China(No.30600693)the Qianjiang Talent Project of Zhejiang Province(No.2010R10068)the Hong Kong Polytechnic University(No.J-BB7P),China
文摘Retinoic acid level in the retina/choroid is altered in induced myopia models.All-trans-retinol dehydrogenase(RDH8) is an important enzyme of retinoic acid metabolism.This study aimed to investigate the association of the RDH8 gene with high myopia.Three single nucleotide polymorphisms(SNPs) [RDH851(rs2233789) ,RDH8E5a(rs1644731) ,and RDH855b(rs3760753) ]were selected,based on the linkage disequilibrium pattern of RDH8 from a previous study,and genotyped for 160 Han Chinese nuclear families with highly myopic(-10 diopters or worse) offspring as well as in an independent group with 166 highly myopic cases(-10 diopters or worse) and 211 controls. Family-based association analysis was performed using the family-based association test(FBAT) package,and genotype relative risk(GRR) was calculated using the GenAssoc program.Population-based association analysis was performed using Chi-square test.These SNPs were in linkage equilibrium with each other.SNPs RDH851(rs2233789) and RDH8E5a(rs1644731) both did not show association with high myopia.SNP RDH855b(rs3760753) demonstrated significant association(P=0.0269) with a GRR of 0.543(95%confidence interval=0.304-0.968,P=0.038) .The association became statistically insignificant,however,after multiple comparison correction.Haplotype analysis did not show a significant association either.Population-based association analysis also showed no significant association(P>0.05) .Our family-and population-based data both suggest that the RDH8 gene is unlikely to be associated with high myopia in Chinese.
