The genomic composition of 1911289, a wheat ( Tritium aestivum L.) maintainer of K-CMS, was examined by several methods, such as genomic in situ hybridization (GISH), biochemical marking, and DNA molecular marking. Th...The genomic composition of 1911289, a wheat ( Tritium aestivum L.) maintainer of K-CMS, was examined by several methods, such as genomic in situ hybridization (GISH), biochemical marking, and DNA molecular marking. The results got by GISH and PCR amplification of dispersed rye-specific repetitive DNA sequence suggested that the alien chromatin in T911289 derived from rye. Specifically PCR amplification of the rye-specific microsatellite primers (SCM9) and seed storage protein analysis indicated that the alien chromatin in T911289 had developed from the short arm of 1R chromosome of rye (1RS). PCR amplification by using microsatellite primers locating on 1BS and seed storage protein analysis also revealed that 1911289 had lost the arm of 1BS or a small distal segment of it. We conclude that T911289 is a heterogeneous population which displays two distinct different types of translocation, i.e. the Robertsonian translocation and small segment translocation. The Robertsonian translocation type observed in our study is different from the 1BL/1RS translocation which is widely used in wheat production; it may be a novel and complex translocation form. Though the linkage between the desirable agronomic traits and the deleterious genes expressed as sticky dough has not got broken in T911289, the recovery of small segment translocation will still benefit the genetic study of wheat and rye.展开更多
Here, we report a method that uses gold nanoparticles (AuNPs) to enhance the specificity of DNA hybridization without reducing its detection sensitivity. The conventional stringent wash method utilizes high-temperat...Here, we report a method that uses gold nanoparticles (AuNPs) to enhance the specificity of DNA hybridization without reducing its detection sensitivity. The conventional stringent wash method utilizes high-temperatureflow-salt conditions to enhance the specificity of DNA hybridization-based assays. This method creates a destabilizing environment for base pairing that affects specific and nonspecific duplexes. Therefore, specificity is achieved at the expense of signal intensity or sensitivity. However, in the proposed wash method, AuNPs predominantly destabilize nonspecific duplexes, offering specificity without compromising sensitivity. This AuNP wash technique has proven to be effective in detecting single nucleotide polymorphisms (SNPs) in genomic samples even at room temperature in a CD-like NanoBioArray (CD-NBA) chip. This method is also robust with sequence variation and is compatible with multiplex DNA analyses on microarrays. Thus, the AuNP wash method could potentially be useful for improving the accuracy of DNA hybridization results.展开更多
Conventional rice breeding has long focused on exploiting the DNA sequence diversity.However,epigenetic diversity,reflected particularly in DNA methylation,can also contribute to phenotypic variation and should not be...Conventional rice breeding has long focused on exploiting the DNA sequence diversity.However,epigenetic diversity,reflected particularly in DNA methylation,can also contribute to phenotypic variation and should not be overlooked in rice breeding.In this study,20 parental lines of indica rice,which are widely used in hybrid rice breeding in China,were analyzed to investigate variations of DNA methylation and its inheritance.The results revealed a wide diversity in DNA methylation among these breeding lines.A positive correlation was seen between DNA methylation and genetic diversity.Furthermore,some of the methylated DNA was inherited in the subsequent generation,regardless of whether they were produced by selfing or hybrid-crossing.This study provides insight into the methylation patterns in rice,and suggests the importance of epigenetic diversity in rice breeding.展开更多
Nanopore has been developed to be a powerful,single-molecule analytical tool for sensing ions,small organic molecules and biomacromolecules such as proteins and DNAs.Generally,the identity of the analyte can be reveal...Nanopore has been developed to be a powerful,single-molecule analytical tool for sensing ions,small organic molecules and biomacromolecules such as proteins and DNAs.Generally,the identity of the analyte can be revealed by current amplitude changes and mean dwell time of the analyte binding events.In some cases,generation of highly characteristic current events affords an alternative way of analyte determination with high confidence level.However,we found that secondary structures in DNA/RNA hybrids might severely hinder the generation of signature events during their translocation through?-hemolysin nanopore.In this report,we propose a strategy to add a certain concentration of urea in the buffer solution for single channel recordings and validate that low concentration of urea can effectively denature the secondary structures in DNA hybrids and recover the generation of signature events.This finding might be useful in other secondary structure-related nanopore sensing activities.展开更多
A versatile DNA recognition system using cadmium sulfide nanoparticles (CdS NPs) as labels was proposed for ultrasensitive detection of specific sequence DNA based on target recycling. This strategy utilized the magne...A versatile DNA recognition system using cadmium sulfide nanoparticles (CdS NPs) as labels was proposed for ultrasensitive detection of specific sequence DNA based on target recycling. This strategy utilized the magnetic particles (MNPs) for the immobilization of linker DNA and CdS NPs and the subsequent target DNA hybridization. Using the unique characteristic of nicking endonuclease for cutting one specific strand of double strand DNA (ds DNA), the linker DNA could be transected and the target DNA could be liberated for re-hybridization and hence the amount of released CdS NPs was enhanced. Due to the advantage of the MNPs and signal amplification from the target recycling, the analyte DNA could be detected by the square-wave stripping voltammetry (SWV) in a wide linear range from 0.4 fM to 100 fM with the detection limit down to 0.08 fM. The proposed DNA detection strategy possesses high sensitivity, satisfactory reproducibility and excellent stability, which might have potential in other DNA biological assays.展开更多
文摘The genomic composition of 1911289, a wheat ( Tritium aestivum L.) maintainer of K-CMS, was examined by several methods, such as genomic in situ hybridization (GISH), biochemical marking, and DNA molecular marking. The results got by GISH and PCR amplification of dispersed rye-specific repetitive DNA sequence suggested that the alien chromatin in T911289 derived from rye. Specifically PCR amplification of the rye-specific microsatellite primers (SCM9) and seed storage protein analysis indicated that the alien chromatin in T911289 had developed from the short arm of 1R chromosome of rye (1RS). PCR amplification by using microsatellite primers locating on 1BS and seed storage protein analysis also revealed that 1911289 had lost the arm of 1BS or a small distal segment of it. We conclude that T911289 is a heterogeneous population which displays two distinct different types of translocation, i.e. the Robertsonian translocation and small segment translocation. The Robertsonian translocation type observed in our study is different from the 1BL/1RS translocation which is widely used in wheat production; it may be a novel and complex translocation form. Though the linkage between the desirable agronomic traits and the deleterious genes expressed as sticky dough has not got broken in T911289, the recovery of small segment translocation will still benefit the genetic study of wheat and rye.
文摘Here, we report a method that uses gold nanoparticles (AuNPs) to enhance the specificity of DNA hybridization without reducing its detection sensitivity. The conventional stringent wash method utilizes high-temperatureflow-salt conditions to enhance the specificity of DNA hybridization-based assays. This method creates a destabilizing environment for base pairing that affects specific and nonspecific duplexes. Therefore, specificity is achieved at the expense of signal intensity or sensitivity. However, in the proposed wash method, AuNPs predominantly destabilize nonspecific duplexes, offering specificity without compromising sensitivity. This AuNP wash technique has proven to be effective in detecting single nucleotide polymorphisms (SNPs) in genomic samples even at room temperature in a CD-like NanoBioArray (CD-NBA) chip. This method is also robust with sequence variation and is compatible with multiplex DNA analyses on microarrays. Thus, the AuNP wash method could potentially be useful for improving the accuracy of DNA hybridization results.
基金supported by the National Natural Science Foundation of China(31071379)the Post-Doctoral Foundation of China(20090450616)a grant from"Yellow Crane"Special Talent Program of Wuhan
文摘Conventional rice breeding has long focused on exploiting the DNA sequence diversity.However,epigenetic diversity,reflected particularly in DNA methylation,can also contribute to phenotypic variation and should not be overlooked in rice breeding.In this study,20 parental lines of indica rice,which are widely used in hybrid rice breeding in China,were analyzed to investigate variations of DNA methylation and its inheritance.The results revealed a wide diversity in DNA methylation among these breeding lines.A positive correlation was seen between DNA methylation and genetic diversity.Furthermore,some of the methylated DNA was inherited in the subsequent generation,regardless of whether they were produced by selfing or hybrid-crossing.This study provides insight into the methylation patterns in rice,and suggests the importance of epigenetic diversity in rice breeding.
基金the National Basic Research Program of China (2013CB932800)the National Natural Science Foundation of China (21175135, 21375130, 21205119, 21475132)the CAS Hundred Talents Program
文摘Nanopore has been developed to be a powerful,single-molecule analytical tool for sensing ions,small organic molecules and biomacromolecules such as proteins and DNAs.Generally,the identity of the analyte can be revealed by current amplitude changes and mean dwell time of the analyte binding events.In some cases,generation of highly characteristic current events affords an alternative way of analyte determination with high confidence level.However,we found that secondary structures in DNA/RNA hybrids might severely hinder the generation of signature events during their translocation through?-hemolysin nanopore.In this report,we propose a strategy to add a certain concentration of urea in the buffer solution for single channel recordings and validate that low concentration of urea can effectively denature the secondary structures in DNA hybrids and recover the generation of signature events.This finding might be useful in other secondary structure-related nanopore sensing activities.
基金supported by the National Natural Science Foundation of China (21025522 & 20890021)the National Natural Science Foundation of China for Creative Research Groups (20821063)the National Basic Research Program of China (2007CB936404)
文摘A versatile DNA recognition system using cadmium sulfide nanoparticles (CdS NPs) as labels was proposed for ultrasensitive detection of specific sequence DNA based on target recycling. This strategy utilized the magnetic particles (MNPs) for the immobilization of linker DNA and CdS NPs and the subsequent target DNA hybridization. Using the unique characteristic of nicking endonuclease for cutting one specific strand of double strand DNA (ds DNA), the linker DNA could be transected and the target DNA could be liberated for re-hybridization and hence the amount of released CdS NPs was enhanced. Due to the advantage of the MNPs and signal amplification from the target recycling, the analyte DNA could be detected by the square-wave stripping voltammetry (SWV) in a wide linear range from 0.4 fM to 100 fM with the detection limit down to 0.08 fM. The proposed DNA detection strategy possesses high sensitivity, satisfactory reproducibility and excellent stability, which might have potential in other DNA biological assays.