scholarly journals Number and size of human X chromosome fragments transferred to mouse cells by chromosome-mediated gene transfer.

1981 ◽  
Vol 1 (5) ◽  
pp. 439-448 ◽  
Author(s):  
A S Olsen ◽  
O W McBride ◽  
D E Moore
Keyword(s):  
Gene Transfer ◽  
X Chromosome ◽  
Deoxyribonucleic Acid ◽  
Single Copy ◽  
Mouse Cell ◽  
Unique Sequence ◽  
Chromosome Fragments ◽  
Mouse Cells ◽  
Chromosomal Sequences ◽  
Kinetics Of

Labeled probes of unique-sequence human X chromosomal deoxyribonucleic acid, prepared by two different procedures, were used to measure the amount of human X chromosomal deoxyribonucleic acid in 12 mouse cell lines expressing human hypoxanthine phosphoribosyltransferase after chromosome-mediated gene transfer. The amount of X chromosomal deoxyribonucleic acid detected by this procedure ranged from undetectable levels in the three stable transformants and some unstable transformants examined to about 20% of the human X chromosome in two unstable transformants. Reassociation kinetics of the X chromosomal probe with deoxyribonucleic acid from the two unstable transformants containing 15 to 20% of the human X chromosome indicate that a single copy of these sequences is present. In one of these lines, the X chromosomal sequences exist as multiple fragments which were not concordantly segregated when the cells were selected for loss of hprt.

Number and size of human X chromosome fragments transferred to mouse cells by chromosome-mediated gene transfer

1981 ◽  
Vol 1 (5) ◽  
pp. 439-448
Author(s):  
A S Olsen ◽  
O W McBride ◽  
D E Moore
Keyword(s):  
Gene Transfer ◽  
X Chromosome ◽  
Deoxyribonucleic Acid ◽  
Single Copy ◽  
Mouse Cell ◽  
Unique Sequence ◽  
Chromosome Fragments ◽  
Mouse Cells ◽  
Chromosomal Sequences ◽  
Kinetics Of

Labeled probes of unique-sequence human X chromosomal deoxyribonucleic acid, prepared by two different procedures, were used to measure the amount of human X chromosomal deoxyribonucleic acid in 12 mouse cell lines expressing human hypoxanthine phosphoribosyltransferase after chromosome-mediated gene transfer. The amount of X chromosomal deoxyribonucleic acid detected by this procedure ranged from undetectable levels in the three stable transformants and some unstable transformants examined to about 20% of the human X chromosome in two unstable transformants. Reassociation kinetics of the X chromosomal probe with deoxyribonucleic acid from the two unstable transformants containing 15 to 20% of the human X chromosome indicate that a single copy of these sequences is present. In one of these lines, the X chromosomal sequences exist as multiple fragments which were not concordantly segregated when the cells were selected for loss of hprt.

IDENTIFICATION AND ISOLATION OF ANTIGENICALLY MARKED COLONIES IN MOUSE CELL CULTURES: A TECHNIQUE FOR DETECTING GENETIC CHANGES

1974 ◽  
Vol 16 (3) ◽  
pp. 565-570
Author(s):  
Ian R. Glen
Keyword(s):  
Gene Transfer ◽  
Cell Cultures ◽  
Inbred Mouse ◽  
Mouse Cell ◽  
Mouse Strains ◽  
Inbred Mouse Strains ◽  
Cell Hybridization ◽  
Genetic Changes ◽  
Pure Cultures ◽  
Mouse Cells

This paper describes a technique by which specific genetic changes involving histocompatability loci in cultured mouse cells can be detected and the altered cells isolated. Reconstruction experiments were carried out using cells from two inbred mouse strains in mixed cultures. These cells were antigenically different and fluorescent antibodies were used to identify cells expressing the desired phenotype in a partially cloned culture. Fluorescing colonies were isolated, recloned, and established as pure cultures. The application of this technique to gene transfer, mutation, and cell hybridization studies is discussed briefly.

scholarly journals Molecular cloning, expression, and chromosomal localization of a human gene encoding the CD33 myeloid differentiation antigen

Blood ◽  
1988 ◽  
Vol 72 (1) ◽  
pp. 314-321
Author(s):  
SC Peiper ◽  
RA Ashmun ◽  
AT Look
Keyword(s):  
Gene Transfer ◽  
Cell Lines ◽  
Cell Sorting ◽  
Human Gene ◽  
Mouse Cell ◽  
Unique Sequence ◽  
Myelogenous Leukemia ◽  
Chromosome 19 ◽  
Gene Encoding ◽  
Sequence Probe

Monoclonal antibodies of the CD33 cluster group recognize a 67- kilodalton (Kd) protein, designated p67, expressed on the surface of normal human myeloid progenitors and leukemic cells from most patients with acute myelogenous leukemia. The human gene encoding p67 was isolated in a mouse genetic background after DNA-mediated gene transfer and fluorescence-activated cell sorting (FACS) for transformants that bound the monoclonal antibody MY9. After three serial rounds of gene transfer and cell sorting, multiple independently derived tertiary mouse cell transformants were obtained that expressed p67. Southern blot analysis revealed that these transformants shared restriction fragments containing highly reiterated human DNA sequences. Two shared EcoRI fragments of 3.3-kilobase (kb) and 9.5-kb pairs were molecularly cloned into bacteriophage vectors. A subsegment of the 3.3-kb fragment lacking repeated sequences was then used as a unique sequence probe to isolate two independent cosmid clones. Cells transfected with DNA from both cosmid clones bound MY9, and the human p67 protein was demonstrated by immunoprecipitation. NFS mice inoculated with a mouse cell transformant coexpressing p67 and the v-fms oncogene product produced antisera that specifically immunoprecipitated p67 from human leukemic cell lines, mouse cell transformants, and mouse cells transfected with the biologically active cosmid clones. The human p67 locus was previously assigned to chromosome 19 by screening a panel of rodent X human somatic cell hybrids with the unique sequence probe. The gene was sublocalized to the q13.3 region of chromosome 19 by in situ hybridization. RNA transcripts of approximately 1.6 kb and 1.4 kb were identified in polyadenylated RNA from human myeloid leukemia cell lines using a probe from the genomic locus. Manipulation of the cloned p67 gene may provide insight into the function of its product and mechanisms regulating its expression.

scholarly journals Transcriptional kinetics of X-chromosome upregulation

2019 ◽  
Author(s):  
Anton J. M. Larsson ◽  
Christos Coucoravas ◽  
Rickard Sandberg ◽  
Björn Reinius
Keyword(s):  
Single Cell ◽  
Rna Sequencing ◽  
X Chromosome ◽  
Single Cell Rna Sequencing ◽  
Allele Specific ◽  
Mouse Cells ◽  
Kinetics Of

Ohno’s hypothesis postulates that X-chromosome upregulation rectifies X-dose imbalance relative to autosomal genes, present in two active copies per cell. Here we dissected X-upregulation into kinetics of transcription, inferred from allele-specific single-cell RNA-sequencing (scRNAseq) data from somatic mouse cells. We confirmed increased X-chromosome expression, and remarkably found that the X-chromosome achieved upregulation by elevated burst frequencies. This provides mechanistic insights into X-chromosome upregulation.

scholarly journals Molecular cloning, expression, and chromosomal localization of a human gene encoding the CD33 myeloid differentiation antigen

Blood ◽  
1988 ◽  
Vol 72 (1) ◽  
pp. 314-321 ◽  
Author(s):  
SC Peiper ◽  
RA Ashmun ◽  
AT Look
Keyword(s):  
Gene Transfer ◽  
Cell Lines ◽  
Cell Sorting ◽  
Human Gene ◽  
Mouse Cell ◽  
Unique Sequence ◽  
Myelogenous Leukemia ◽  
Chromosome 19 ◽  
Gene Encoding ◽  
Sequence Probe

Abstract Monoclonal antibodies of the CD33 cluster group recognize a 67- kilodalton (Kd) protein, designated p67, expressed on the surface of normal human myeloid progenitors and leukemic cells from most patients with acute myelogenous leukemia. The human gene encoding p67 was isolated in a mouse genetic background after DNA-mediated gene transfer and fluorescence-activated cell sorting (FACS) for transformants that bound the monoclonal antibody MY9. After three serial rounds of gene transfer and cell sorting, multiple independently derived tertiary mouse cell transformants were obtained that expressed p67. Southern blot analysis revealed that these transformants shared restriction fragments containing highly reiterated human DNA sequences. Two shared EcoRI fragments of 3.3-kilobase (kb) and 9.5-kb pairs were molecularly cloned into bacteriophage vectors. A subsegment of the 3.3-kb fragment lacking repeated sequences was then used as a unique sequence probe to isolate two independent cosmid clones. Cells transfected with DNA from both cosmid clones bound MY9, and the human p67 protein was demonstrated by immunoprecipitation. NFS mice inoculated with a mouse cell transformant coexpressing p67 and the v-fms oncogene product produced antisera that specifically immunoprecipitated p67 from human leukemic cell lines, mouse cell transformants, and mouse cells transfected with the biologically active cosmid clones. The human p67 locus was previously assigned to chromosome 19 by screening a panel of rodent X human somatic cell hybrids with the unique sequence probe. The gene was sublocalized to the q13.3 region of chromosome 19 by in situ hybridization. RNA transcripts of approximately 1.6 kb and 1.4 kb were identified in polyadenylated RNA from human myeloid leukemia cell lines using a probe from the genomic locus. Manipulation of the cloned p67 gene may provide insight into the function of its product and mechanisms regulating its expression.

Generation of sequence-tagged sites from Xp22.3 by isolating commonAlu-PCR products of radiation hybrids retaining overlapping human X chromosome fragments

1996 ◽  
Vol 97 (5) ◽  
pp. 604-610 ◽  
Author(s):  
I. A. Glass ◽  
M. Passage ◽  
L. Bernatowicz ◽  
E. C. Salido ◽  
T. Mohandas ◽  
...  
Keyword(s):  
X Chromosome ◽  
Sequence Tagged Sites ◽  
Radiation Hybrids ◽  
Pcr Products ◽  
Chromosome Fragments

scholarly journals Cloning and characterization of small circular DNA from Chinese hamster ovary cells.

1984 ◽  
Vol 4 (1) ◽  
pp. 173-180 ◽  
Author(s):  
S W Stanfield ◽  
D R Helinski
Keyword(s):  
Chinese Hamster Ovary ◽  
Cho Cells ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Single Copy ◽  
Chromosomal Dna ◽  
Cho Cell ◽  
Ovary Cells ◽  
Chromosomal Sequences

Small polydisperse circular (spc) DNA was isolated and cloned, using BglII from Chinese hamster ovary (CHO) cells. The properties of 47 clones containing at least 43 different BglII fragments are reported. The majority of the clones probably contain entire sequences from individual spcDNA molecules. Most of the clones were homologous to sequences in CHO cell chromosomal DNA, and many were also homologous to mouse LMTK- cell chromosomal sequences. The majority of homologous CHO cell chromosomal sequences were repetitive, although a few may be single copy. Only a small fraction of cloned spcDNA molecules were present in every cell; most occurred less frequently than once in 15 cells. Localization studies indicated that at least a portion of spcDNA is associated with the nucleus in CHO cells.

Differentiation of two mouse cell lines is associated with hypomethylation of their genomes

1984 ◽  
Vol 4 (9) ◽  
pp. 1800-1806
Author(s):  
T H Bestor ◽  
S B Hellewell ◽  
V M Ingram
Keyword(s):  
Dna Methyltransferase ◽  
Cell Types ◽  
Mouse Cell ◽  
F9 Cells ◽  
Dna Restriction Fragments ◽  
Dna Restriction ◽  
F9 Embryonal Carcinoma Cells ◽  
Murine Erythroleukemia ◽  
Kinetics Of

Methyl-accepting assays and a sensitive method for labeling specific CpG sites have been used to show that the DNA of F9 embryonal carcinoma cells decreases in 5-methylcytosine content by ca. 9% during retinoic acid-induced differentiation, whereas the DNA of dimethyl sulfoxide-induced Friend murine erythroleukemia (MEL) cells loses ca. 3.8% of its methyl groups. These values correspond to the demethylation of 2.2 X 10(6) and 0.9 X 10(6) 5'-CpG-3' sites per haploid genome in differentiating F9 and MEL cells, respectively. Fluorography of DNA restriction fragments methylated in vitro and displayed on agarose gels showed that demethylation occurred throughout the genome. In uninduced F9 cells, the sequence TCGA tended to be more heavily methylated than did the sequence CCGG, whereas this tendency was reversed in MEL cells. The kinetics of in vitro DNA methylation reactions catalyzed by MEL cell DNA methyltransferase showed that substantial numbers of hemimethylated sites accumulate in the DNA of terminally differentiating F9 and MEL cells, implying that a partial loss of DNA-methylating activity may accompany terminal differentiation in these two cell types.

scholarly journals Involvement of plastid, mitochondrial and nuclear genomes in plant-to-plant horizontal gene transfer

2014 ◽  
Vol 83 (4) ◽  
pp. 317-323 ◽  
Author(s):  
Maria Virginia Sanchez-Puerta
Keyword(s):  
Gene Transfer ◽  
Horizontal Gene Transfer ◽  
Plant Mitochondria ◽  
Genetic Material ◽  
Single Copy ◽  
Convincing Evidence ◽  
Cell Contact ◽  
Plant Mtdna ◽  
Nuclear Genomes ◽  
Foreign Genes

This review focuses on plant-to-plant horizontal gene transfer (HGT) involving the three DNA-containing cellular compartments. It highlights the great incidence of HGT in the mitochondrial genome (mtDNA) of angiosperms, the increasing number of examples in plant nuclear genomes, and the lack of any convincing evidence for HGT in the well-studied plastid genome of land plants. Most of the foreign mitochondrial genes are non-functional, generally found as pseudogenes in the recipient plant mtDNA that maintains its functional native genes. The few exceptions involve chimeric HGT, in which foreign and native copies recombine leading to a functional and single copy of the gene. Maintenance of foreign genes in plant mitochondria is probably the result of genetic drift, but a possible evolutionary advantage may be conferred through the generation of genetic diversity by gene conversion between native and foreign copies. Conversely, a few cases of nuclear HGT in plants involve functional transfers of novel genes that resulted in adaptive evolution. Direct cell-to-cell contact between plants (e.g. host-parasite relationships or natural grafting) facilitate the exchange of genetic material, in which HGT has been reported for both nuclear and mitochondrial genomes, and in the form of genomic DNA, instead of RNA. A thorough review of the literature indicates that HGT in mitochondrial and nuclear genomes of angiosperms is much more frequent than previously expected and that the evolutionary impact and mechanisms underlying plant-to-plant HGT remain to be uncovered.

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