The distribution of repetitive DNAs along chromosomes is one of the crucial elements for understanding the organization and the evolution of plant genomes. Using a modified genomic in situ hybridization （GISH） proce...The distribution of repetitive DNAs along chromosomes is one of the crucial elements for understanding the organization and the evolution of plant genomes. Using a modified genomic in situ hybridization （GISH） procedure, fluorescence in situ hybridization （FISH） with genomic DNA to their own chromosomes （called self-genomic in situ hybridization, self-GISH） was carried out in six selected plant species with different genome size and amount of repetitive DNA. Nonuniform distribution of the fluorescent labeled probe DNA was observed on the chromosomes of all the species that were tested. The signal patterns varied among species and were related to the genome size. The chromosomes of the small Arabidopsis genome were labeled almost only in the pericentromeric regions and the nucleolus organizer regions （NORs）. The signals in the relatively small genomes, rice, sorghum, and Brassica oleracea var. capitata L., were dispersed along the chromosome lengths, with a predominant distribution in the pericentromeric or proximal regions and some heterochromatic arms. All chromosomes of the large genomes, maize and barley, were densely labeled with strongly labeled regions and weakly labeled or unlabeled regions being arranged alternatively throughout the lengths. In addition, enhanced signal bands were shown in all pericentromeres and the NORs in B. oleracea var. capitata, and in all pericentromeric regions and certain intercalary sites in barley. The enhanced signal band pattern in barley was found consistent with the N-banding pattern of this species. The GISH with self-genomic DNA was compared with FISH with Cot-1 DNA in rice, and their signal patterns are found to be basically consistent. Our results showed that the self-GISH signals actually reflected the hybridization of genomic repetitive DNAs to the chromosomes, thus the self-GISH technique would be useful for revealing the distribution of the regions where repetitive DNAs concentrate along chromosomes and some chromatin differentiation associated with repetitive DNAs in plants.展开更多
Double helix DNAs become intertwined around one another during replication and recombination.Here we used magnetic tweezers to make braided DNA molecules and measured their torques under various catenations(Ca)at forc...Double helix DNAs become intertwined around one another during replication and recombination.Here we used magnetic tweezers to make braided DNA molecules and measured their torques under various catenations(Ca)at forces ranging from 0.3 to 8 pN.Images of braided DNA constructs under tensions were captured by scanning electron microscopy which showed major and minor grooves of DNAs and plectonemes of the braids.When the two DNA molecules were braided,the extension decreased as the catenation increased from 0 to 50 turns.We used a thermodynamic Maxwell relation to deduce the torque by integrating the change in the braid extension as a function of the force.The torque increased with the catenation,force and intertether distance until the catenation reached a buckling point.Under the condition of 2 pN force and Ca=20,the torque was computed to be 31,21 and 15 pN nm for the braids of which the intertether distances were 54%,31%and 26%of the DNA contour length,respectively.At an 8.03 pN holding force,the torque was computed to be 76 pN nm as the catenation increased from 0 to 30 turns,or as the catenation density varied from 0 to 0.053.The torque reached a plateau when the catenation increased above 20,indicating formation of braid-plectonemes.The twist modulus increased with the catenation prior to reaching a peak.Before reaching the peak,the moduli were higher than those of a single twisted DNA under the same catenation and applied force.Our experimental data agrees well with the calculation results by a recently developed semiflexible polymer model.Our measurements of the nonlinear torque of the braid establish new fundamental properties of DNA intertwining,which is key to understanding DNA replication and gene expression.The speaker will also introduce briefly other projects in the Xiao group including direct measurements of theforce spectrum of single unlabeled proteins such as adhesive nano-fibers for biofilm,the screening of integrin-targeted peptides drugs by single cell approaches,and the micromechanical approach for determining the survival rate of stem cells.展开更多
基金This work was supported by the National Natural Sciences Foundation of China (No. 39870423).
文摘The distribution of repetitive DNAs along chromosomes is one of the crucial elements for understanding the organization and the evolution of plant genomes. Using a modified genomic in situ hybridization （GISH） procedure, fluorescence in situ hybridization （FISH） with genomic DNA to their own chromosomes （called self-genomic in situ hybridization, self-GISH） was carried out in six selected plant species with different genome size and amount of repetitive DNA. Nonuniform distribution of the fluorescent labeled probe DNA was observed on the chromosomes of all the species that were tested. The signal patterns varied among species and were related to the genome size. The chromosomes of the small Arabidopsis genome were labeled almost only in the pericentromeric regions and the nucleolus organizer regions （NORs）. The signals in the relatively small genomes, rice, sorghum, and Brassica oleracea var. capitata L., were dispersed along the chromosome lengths, with a predominant distribution in the pericentromeric or proximal regions and some heterochromatic arms. All chromosomes of the large genomes, maize and barley, were densely labeled with strongly labeled regions and weakly labeled or unlabeled regions being arranged alternatively throughout the lengths. In addition, enhanced signal bands were shown in all pericentromeres and the NORs in B. oleracea var. capitata, and in all pericentromeric regions and certain intercalary sites in barley. The enhanced signal band pattern in barley was found consistent with the N-banding pattern of this species. The GISH with self-genomic DNA was compared with FISH with Cot-1 DNA in rice, and their signal patterns are found to be basically consistent. Our results showed that the self-GISH signals actually reflected the hybridization of genomic repetitive DNAs to the chromosomes, thus the self-GISH technique would be useful for revealing the distribution of the regions where repetitive DNAs concentrate along chromosomes and some chromatin differentiation associated with repetitive DNAs in plants.
基金supported by the National Science Foundation of China ( 11772133, 11372116)the Fundamental Research Funds for the Central Universities ( HUST 0118012051)supported by the NIH through grants ( R01-GM105847,U54-CA193419)
文摘Double helix DNAs become intertwined around one another during replication and recombination.Here we used magnetic tweezers to make braided DNA molecules and measured their torques under various catenations(Ca)at forces ranging from 0.3 to 8 pN.Images of braided DNA constructs under tensions were captured by scanning electron microscopy which showed major and minor grooves of DNAs and plectonemes of the braids.When the two DNA molecules were braided,the extension decreased as the catenation increased from 0 to 50 turns.We used a thermodynamic Maxwell relation to deduce the torque by integrating the change in the braid extension as a function of the force.The torque increased with the catenation,force and intertether distance until the catenation reached a buckling point.Under the condition of 2 pN force and Ca=20,the torque was computed to be 31,21 and 15 pN nm for the braids of which the intertether distances were 54%,31%and 26%of the DNA contour length,respectively.At an 8.03 pN holding force,the torque was computed to be 76 pN nm as the catenation increased from 0 to 30 turns,or as the catenation density varied from 0 to 0.053.The torque reached a plateau when the catenation increased above 20,indicating formation of braid-plectonemes.The twist modulus increased with the catenation prior to reaching a peak.Before reaching the peak,the moduli were higher than those of a single twisted DNA under the same catenation and applied force.Our experimental data agrees well with the calculation results by a recently developed semiflexible polymer model.Our measurements of the nonlinear torque of the braid establish new fundamental properties of DNA intertwining,which is key to understanding DNA replication and gene expression.The speaker will also introduce briefly other projects in the Xiao group including direct measurements of theforce spectrum of single unlabeled proteins such as adhesive nano-fibers for biofilm,the screening of integrin-targeted peptides drugs by single cell approaches,and the micromechanical approach for determining the survival rate of stem cells.
