Precise localization of human beta-globin gene complex on chromosome 11.

Gusella, James F.; Varsanyi-Breiner, Avia; Kao, Fa‐Ten; Jones, Carol; Puck, Theodore T.; Keys, Cheryl; Orkin, Stuart H.; Housman, David E.

doi:10.1073/pnas.76.10.5239

Cited by 146 publications

(75 citation statements)

References 27 publications

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“…For these experiments, we needed a probe that would be specific for detecting small amounts of human DNA in a background of a large excess of Chinese hamster DNA. It had previously been demonstrated that human middle repetitive DNA sequences, which are distributed throughout the human genome every 2 to 3 kilobases, do not cross-hybridize to any significant extent with the analogous sequences from the Chinese hamster genome (14,15,23). Therefore, human middle repetitive sequences appeared to be the distinguishing probe we needed.…”

Section: Methodsmentioning

confidence: 99%

“…These libraries will be screened by using the Benton and Davis plaque hybridization technique with total nick-translated human DNA as a probe (4). As described by Gusella et al, this procedure distinguishes phage recombinants containing human DNA from recombinants containing only rodent DNA, again by virtue of differential hybridization of human and rodent middle repetitive DNA sequences (14,15). Since middle repetitive sequences are dispersed in an apparently random manner every 2 to 3 kilobases through the human genome, virtually every phage recombinant (with inserts of -20 kilobases) that contains unique, single-copy human sequences will most likely also contain at least one middle repetitive sequence and will be identified in the screening process (23).…”

Section: Methodsmentioning

confidence: 99%

“…These dot blots were hybridized to total genomic human DNA that had been labeled with 32P by nick translation and then washed and exposed to X-ray film as also described above. In preparations of total nicktranslated human DNA, the only sequences which are present in a high enough copy number to give a detectable hybridization signal are the highly repeated sequences (15,16). An autoradiograph of one such blot is shown in Fig.…”

Section: Methodsmentioning

confidence: 99%

“…Using this procedure, we focused on transferring the human genes encoding leucyl-and asparaginyl-tRNA synthetase into the appropriate CHO cell mutants because the presence of human DNA can be monitored against the background of rodent DNA by cytological as well as nucleic acid hybridization procedures (10,14,15,22). We have previously isolated interspecific somatic cell hybrids between human leukocytes and CHO cell mutants with temperature-sensitive leucyl-or asparaginyl-tRNA synthetases, which contain and express the corresponding human aminoacyl tRNA-synthetase genes (6,8,9).…”

Section: Transfer Of Human Genes Into Cho Cell Mutants 893mentioning

confidence: 99%

“…Since these hybrids contained very few human chromosomes to begin with, they represented a significant enrichment of the human genes of interest. Hybrid cell lines were subjected to high doses of y-irradiation to extensively fragment the chromosomes and were refused immediately to the original temperature-sensitive CHO mutant, and "secondary" hybrids were isolated at 39°C to require (14,15), which will provide at least a 1,000-fold purification of the genes of interest without the use of specific probes. The procedures we describe can be applied to enrich for any gene for which a selective system is available.…”

Section: Transfer Of Human Genes Into Cho Cell Mutants 893mentioning

confidence: 99%

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Efficient procedure for transferring specific human genes into Chinese hamster cell mutants: interspecific transfer of the human genes encoding leucyl- and asparaginyl-tRNA synthetases.

Cirullo¹,

Dana²,

Wasmuth³

1983

Mol. Cell. Biol.

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We have developed a simple and efficient procedure for transferring specific human genes into mutant Chinese hamster ovary cell recipients that does not rely on using calcium phosphate-precipitated high-molecular-weight DNA. Interspecific cell hybrids between human leukocytes and temperature-sensitive Chinese hamster cell mutants with either a thermolabile leucyl-tRNA synthetase or a thermolabile asparaginyl-tRNA synthetase were used as the starting material in these experiments. These hybrids contain only one or a few human chromosomes and require expression of the appropriate human aminoacyl-tRNA synthetase gene to grow at 39°C. Hybrids were exposed to very high doses of -y-irradiation to extensively fragment the chromosomes and re-fused immediately to the original temperature-sensitive Chinese hamster mutant, and secondary hybrids were isolated at 39°C. Secondary hybrids, which had retained small fragments of the human genome containing the selected gene, were subjected to another round of irradiation, refusion, and selection at 39°C to reduce the amount of human DNA even further. Using this procedure, we have constructed Chinese hamster cell lines that express the human genes encoding either asparaginyl-or leucyl-tRNA synthetase, yet less than 0.1% of their DNA is derived from the human genome, as quantitated by a sensitive dot-blot nucleic acid hybridization procedure. Analysis of these cell lines with Southern blots confirmed the presence of a small number of restriction endonuclease fragments containing human DNA specifically. These cell lines represent a convenient and simple means to clone the human genomic sequences of interest.The genes encoding various components of the protein synthetic machinery in mammalian cells represent a very large family offunctionally related genes. To begin understanding how the expression of this family of genes is coordinated, we are using a combined biochemical and genetic approach to analyze the complex process of protein synthesis. The large number of different Chinese hamster cell mutants that have been isolated with alterations in various protein synthesis components, including ribosomal proteins and aminoacyl-tRNA synthetases, makes this large family of genes especially suitable for genetic analysis and genetic manipulation (1,3,5,13,28,33,34). In Chinese hamster ovary (CHO) cells, two of these genes, leuS and asnS, which encode leucyl-tRNA synthetase and asparaginyl-tRNA synthetase, respectively, are particularly amenable to detailed genetic studies. Conditionally lethal, temperature-sensitive mutants with alterations in either of these genes can be isolated at very high frequencies (1,29,31). In addition, there is a simple counterselective system, growth at an elevated temperature, to isolate revertants in which the temperaturesensitive phenotype of the mutants has been suppressed by second-site intergenic or intragenic mutations. Because of the availability of large numbers of mutants and revertants, the leuS and asnS genes are most amenable to detailed studies o...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Transfer Of Human Genes Into Cho Cell Mutants 893mentioning

confidence: 99%

Section: Transfer Of Human Genes Into Cho Cell Mutants 893mentioning

confidence: 99%

See 3 more Smart Citations

Efficient procedure for transferring specific human genes into Chinese hamster cell mutants: interspecific transfer of the human genes encoding leucyl- and asparaginyl-tRNA synthetases.

Cirullo¹,

Dana²,

Wasmuth³

1983

Mol. Cell. Biol.

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Genome-wide search for linkage of bipolar affective disorders in a very large pedigree derived from a homogeneous population in Quebec points to a locus of major effect on chromosome 12q23-q24

Villeneuve²,

et al. 1999

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We completed a genome-wide scan for susceptibility loci for bipolar affective disorders in families derived from a rather homogeneous population in the Province of Québec. The genetic homogeneity of this population stems from the migration of founding families into this relatively isolated area of Québec in the 1830s. A possible founder effect, combined with a prevalence of very large families, makes this population ideal for linkage studies. Genealogies for probands can be readily constructed from a population database of acts of baptism and marriage from the early 1830s up to the present time (the BALSAC register). We chose probands with a DSM III diagnosis of bipolar affective disorder and who may be grouped within large families having genealogical origins with the founding population of the Saguenay-Lac-St-Jean area. Living members (n approximately 120) of a very large pedigree were interviewed using the Structured Clinical Interview for DSM III (SCID I), SCID II, and with a family history questionnaire. A diagnostic panel evaluated multisource information (interview, medical records, family history) and pronounced best-estimate consensus diagnoses on all family members. Linkage, SimAPM, SimIBD, and sib-pair analyses have been performed with 332 microsatellite probes covering the entire genome at an average spacing of 11 cM. GENEHUNTER and haplotype analyses were performed on regions of interest. Analysis of a second large pedigree in the same regions of interest permitted confirmation of presumed linkages found in the region of chromosome 12q23-q24.

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Inhibition of Sickle Hemoglobin Polymerization as a Basis for Therapeutic Approaches to Sickle‐Cell Anemia

Noguchi

Schechter

Haley³

et al. 2003

Burger's Medicinal Chemistry and Drug Discovery

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This chapter reviews current understanding of the molecular and cellular basis of the pathophysiology of sickle‐cell anemia, with emphasis on the biophysics of the intracellular polymerization of sickle hemogobin. Factors that modify the severity of the disease manifestations, primarily by altering polymerization tendency, are discussed. This framework is used to present an overview of the major rational approaches to the therapy of sickle‐cell anemia based on inhibiting polymerization with agents that affect either the hemoglobin molecule or the sickle erythrocyte. Discussion of agents that may decrease vascular entrapment of sickle erythrocytes is also presented. Detailed review of the most important current approaches to therapy—pharmacologically elevating fetal hemoglobin—focuses primarily on hydroxyurea, the only drug currently FDA approved for treating sickle‐cell anemia. A brief discussion of other approaches, including stem cell transplantation and gene therapy, concludes the chapter.

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Precise localization of human beta-globin gene complex on chromosome 11.

Cited by 146 publications

References 27 publications

Efficient procedure for transferring specific human genes into Chinese hamster cell mutants: interspecific transfer of the human genes encoding leucyl- and asparaginyl-tRNA synthetases.

Efficient procedure for transferring specific human genes into Chinese hamster cell mutants: interspecific transfer of the human genes encoding leucyl- and asparaginyl-tRNA synthetases.

Genome-wide search for linkage of bipolar affective disorders in a very large pedigree derived from a homogeneous population in Quebec points to a locus of major effect on chromosome 12q23-q24

Inhibition of Sickle Hemoglobin Polymerization as a Basis for Therapeutic Approaches to Sickle‐Cell Anemia

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