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
Comparing to its sister-family (Rhinolophidae), Hipposideridae was less studied by cytogenetic approaches. Only a few high-resolution G-banded karyotypes have been reported so far, and most of the conclusions on the...Comparing to its sister-family (Rhinolophidae), Hipposideridae was less studied by cytogenetic approaches. Only a few high-resolution G-banded karyotypes have been reported so far, and most of the conclusions on the karyotypic evolution in Hipposideridae were based on conventional Giemsa-staining. In this study, we applied comparative chromosome painting, a method of choice for genome-wide comparison at the molecular level, and G- and C-banding to establish comparative map between five hipposiderid species from China, using a whole set of chromosome-specific painting probes from one of them (Aselliscus stoliczkanus). G-band and C-band comparisons between homologous segments defined by chromosome painting revealed that Robertsonian translocations, paracentric inversions and heterochromatin addition could be the main mechanism of chromosome evolution in Hipposideridae. Comparative analysis of the conserved chromosomal segments among five hipposiderid species and outgroup species suggests that bi-armed chromosomes should be included into the ancestral karyotype of Hipposideridae, which was previously believed to be exclusively composed of acrocentric chromosomes.展开更多
Lung cancer is a leading cause of cancer death worldwide. Some lung cancer patients correlate with a gas of radon besides smoking. To search for common chromosomal aberrations in lung cancer cell lines established fro...Lung cancer is a leading cause of cancer death worldwide. Some lung cancer patients correlate with a gas of radon besides smoking. To search for common chromosomal aberrations in lung cancer cell lines established from patients induced by different factors, a combined approach of chromosome sorting, forward and reverse chromosome painting was used to characterize karyotypes of two lung adenocarcinoma cell lines: A549 and GLC-82 with the latter line derived from a patient who has suffered long-term exposure to environmental radon gas pollution. The chromosome painting results revealed that complex chromosomal rearrangements occurred in these two lung adenocarcinoma cell lines. Thirteen and twenty-four abnormal chromosomes were identified An A549 and GLC-82 cell lines, respectively. Almost half of abnormal chromosomes in these two cell lines were formed by non-reciprocal translocations, the others were derived from deletions and duplication/or amplification in some chromosomal regions. Furthermore, two apparently common breakpoints, HSA8q24 and 12q14 were found in these two lung cancer cell lines.展开更多
Sugarcane has recently attracted increasing attention for its potential as a source of sugar and bioethanol,so increasing its yield is essential to ensure the sugar security and bioenergy production.Intergeneric hybri...Sugarcane has recently attracted increasing attention for its potential as a source of sugar and bioethanol,so increasing its yield is essential to ensure the sugar security and bioenergy production.Intergeneric hybridization is a highly efficient method to produce new genetic variants of crop plants,particularly those species with high ploidy such as sugarcane(Saccharum spp.).Tripidium arundinaceum exhibits many desirable agronomic traits,and has been widely studied to produce hybrids with improved stress tolerance and other characteristics in sugarcane breeding.However,the genetic relationship between T.arundinaceum and Saccharum species,and the individual T.arundinaceum chromosomal compositions in sugarcane hybrids are still elusive.Here we used whole-genome single-nucleotide polymorphisms(SNPs)to ascertain the phylogenetic relationships between these species and found that T.arundinaceum is more closely related to Saccharum than Sorghum,in contrast to the previous narrow genetic analyses using chloroplast DNA.Additionally,oligonucleotide(oligo)-based chromosome-specific painting derived from Saccharum officinarum was able to distinctly identify the chromosomes of T.arundinaceum.We developed the oligo-genomic in situ hybridization(GISH)system for the first time,to unveil the novel chromosome translocations and the transmission of individual T.arundinaceum chromosomes in sugarcane progeny.Notably,we discovered that the chromosomal transmission of T.arundinaceum exhibited several different inheritance modes,including n,2n,and over 2n in the BC1 progenies.Such inheritance patterns may have resulted from first division restitution(FDR)or FDR+nondisjunction of a chromosome with the sister chromatids in the second meiosis division/second division restitution(FDR+NSC/SDR)model during meiosis.These results will be of substantial benefit for the further selection of T.arundinaceum chromosomes for sugarcane genetic improvement.展开更多
Since Ldecke first successfully introduced the PCR technique into microdissectionon banded chromosomes, direct construction of a bandspecific DNA probe pool bymicrodissection has been widely used in cytogenetic and mo...Since Ldecke first successfully introduced the PCR technique into microdissectionon banded chromosomes, direct construction of a bandspecific DNA probe pool bymicrodissection has been widely used in cytogenetic and molecular genetic studies.It can be used not only for the detection of chromosome abnormalities at chromosomeband level but also for the generation of chromosome band-specific DNA probes andgenetic makers.展开更多
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%。
基金supported by grants from the National Natural Science Foundation of China(30770293)the Ministry of Science and Technology of China(2005DKA21502)
文摘Comparing to its sister-family (Rhinolophidae), Hipposideridae was less studied by cytogenetic approaches. Only a few high-resolution G-banded karyotypes have been reported so far, and most of the conclusions on the karyotypic evolution in Hipposideridae were based on conventional Giemsa-staining. In this study, we applied comparative chromosome painting, a method of choice for genome-wide comparison at the molecular level, and G- and C-banding to establish comparative map between five hipposiderid species from China, using a whole set of chromosome-specific painting probes from one of them (Aselliscus stoliczkanus). G-band and C-band comparisons between homologous segments defined by chromosome painting revealed that Robertsonian translocations, paracentric inversions and heterochromatin addition could be the main mechanism of chromosome evolution in Hipposideridae. Comparative analysis of the conserved chromosomal segments among five hipposiderid species and outgroup species suggests that bi-armed chromosomes should be included into the ancestral karyotype of Hipposideridae, which was previously believed to be exclusively composed of acrocentric chromosomes.
基金supported partly by grants from the Ministry of Science and Technology of China(2005DKA21502)the Joint Foundation of Science and Technology Bureau of Yunnan Province and Kunming Medical University(2007C0024R)
文摘Lung cancer is a leading cause of cancer death worldwide. Some lung cancer patients correlate with a gas of radon besides smoking. To search for common chromosomal aberrations in lung cancer cell lines established from patients induced by different factors, a combined approach of chromosome sorting, forward and reverse chromosome painting was used to characterize karyotypes of two lung adenocarcinoma cell lines: A549 and GLC-82 with the latter line derived from a patient who has suffered long-term exposure to environmental radon gas pollution. The chromosome painting results revealed that complex chromosomal rearrangements occurred in these two lung adenocarcinoma cell lines. Thirteen and twenty-four abnormal chromosomes were identified An A549 and GLC-82 cell lines, respectively. Almost half of abnormal chromosomes in these two cell lines were formed by non-reciprocal translocations, the others were derived from deletions and duplication/or amplification in some chromosomal regions. Furthermore, two apparently common breakpoints, HSA8q24 and 12q14 were found in these two lung cancer cell lines.
基金funded by the Central Government and Local Science and Technology Development Special Project,China(2022L3086)the Sugarcane Research Foundation of Guangxi University,China(2022GZB006)+3 种基金supported by the National Natural Science Foundation of China(31771863)the Academy of Sugarcane and Sugar Industry,Guangxi University,China(ASSI-2023009)an independent fund of Guangxi Key Laboratory of Sugarcane Biology,China(GXKLSCB-20190201)the China Agriculture Research System of MOF and MARA(CARS-20-1-5)。
文摘Sugarcane has recently attracted increasing attention for its potential as a source of sugar and bioethanol,so increasing its yield is essential to ensure the sugar security and bioenergy production.Intergeneric hybridization is a highly efficient method to produce new genetic variants of crop plants,particularly those species with high ploidy such as sugarcane(Saccharum spp.).Tripidium arundinaceum exhibits many desirable agronomic traits,and has been widely studied to produce hybrids with improved stress tolerance and other characteristics in sugarcane breeding.However,the genetic relationship between T.arundinaceum and Saccharum species,and the individual T.arundinaceum chromosomal compositions in sugarcane hybrids are still elusive.Here we used whole-genome single-nucleotide polymorphisms(SNPs)to ascertain the phylogenetic relationships between these species and found that T.arundinaceum is more closely related to Saccharum than Sorghum,in contrast to the previous narrow genetic analyses using chloroplast DNA.Additionally,oligonucleotide(oligo)-based chromosome-specific painting derived from Saccharum officinarum was able to distinctly identify the chromosomes of T.arundinaceum.We developed the oligo-genomic in situ hybridization(GISH)system for the first time,to unveil the novel chromosome translocations and the transmission of individual T.arundinaceum chromosomes in sugarcane progeny.Notably,we discovered that the chromosomal transmission of T.arundinaceum exhibited several different inheritance modes,including n,2n,and over 2n in the BC1 progenies.Such inheritance patterns may have resulted from first division restitution(FDR)or FDR+nondisjunction of a chromosome with the sister chromatids in the second meiosis division/second division restitution(FDR+NSC/SDR)model during meiosis.These results will be of substantial benefit for the further selection of T.arundinaceum chromosomes for sugarcane genetic improvement.
文摘Since Ldecke first successfully introduced the PCR technique into microdissectionon banded chromosomes, direct construction of a bandspecific DNA probe pool bymicrodissection has been widely used in cytogenetic and molecular genetic studies.It can be used not only for the detection of chromosome abnormalities at chromosomeband level but also for the generation of chromosome band-specific DNA probes andgenetic makers.