Report of the committee on the genetic constitution of chromosomes 13 to 22

Ferguson-Smith, M.A.; Westerveld, A.

doi:10.1159/000131696

Cited by 11 publications

(2 citation statements)

References 6 publications

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“…In the course of our attempts to clone several alleles of D14S1 (6)(7)(8), a recombinant DNA library consisting of large fragments having one BamHI end and one EcoRI end was constructed in the standard bacteriophage X vector Charon 30 (13). Ligated DNA was packaged in vitro and plated on a host strain (DB1170) that is mutant at the recB, recC, and sbcB loci.…”

Section: Resultsmentioning

confidence: 99%

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Propagation of some human DNA sequences in bacteriophage lambda vectors requires mutant Escherichia coli hosts.

Wyman¹,

Wolfe²,

Botstein³

1985

Proc. Natl. Acad. Sci. U.S.A.

108

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The growth of clones of human genomic DNA fragments in a bacteriophage X vector has been examined in a number of different Escherichia coli hosts. A large proportion (8.9%) of the phages carrying different fragments of the human genome fail to grow on standard rec' hosts but will grow on hosts carrying mutations in the recB, recC, and sbcB genes. Heteroduplex analysis in the electron microscope of DNA from four of these phages revealed substantial secondary structure, including snap-back regions 200-500 base pairs in length. Such structures were not found in phages from the same DNA library that grow in rec' hosts. Recombinant DNA libraries in bacteriophage X vectors are generally found to contain most of the sequences of the genomes from which they are derived (1, 2). Nevertheless, anecdotal reports of sequences that cannot be found in libraries of eukaryotic genomes are fairly common. One striking example is the hypervariable region 3' to the a-globin locus, which has resisted strenuous efforts at cloning (S. Goodbourn and S. Orkin, personal communications). Another example is the junction fragment from a translocation between chromosomes 6 and 10 in a murine plasmacytoma (3). A related phenomenon is the observation of discrepancies between cloned and genomic sequences that are the result of deletions occurring during cloning of mammalian DNA (3)(4)(5). Only some of these deletions can be prevented by growth on a recA host.For the past few years we have been studying a highly polymorphic locus in humans, now known as D14S1, which was identified by hybridization of gel transfers of genomic DNA using an adjacent single-copy fragment as probe (6-8).Efforts to obtain clones of the sequences surrounding and including the polymorphic site itself from the Maniatis human genomic library (2) and from libraries made in our laboratory, using conventional cloning conditions as well as hosts carrying mutations in the recA gene, were all unsuccessful. Given the rarity in general of highly polymorphic DNA sequences in the human genome and the striking difficulty in recovering the variable segments in two of the most studied ones (a globin and D14S1), we became concerned that there might be a systematic difficulty in cloning certain human DNA sequences. Further, we speculated that the difficulty might sometimes be associated with a high degree of polymorphism.Recently, Leach and Stahl (9) reported that inverted repetitions cannot be cloned into phage X vectors unless mutant hosts are used. We report here that such a host, which contains mutant recB, recC, and sbcB genes, will propagate many phage X clones containing random human DNA fragments, including D14S1, that cannot be grown on the Escherichia coli hosts commonly used in the selection and propagation of recombinant DNA genome libraries. MATERIALS AND METHODSBacterial Strains. With the exception of strains BNN45 and SF8, all strains used are derivatives of the E. coli K-12 strain AB1157; they have the following markers in common: leu6 aral4 his4 thrl thil lac Y1...

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Section: Resultsmentioning

confidence: 99%

“…For the past few years we have been studying a highly polymorphic locus in humans, now known as D14S1, which was identified by hybridization of gel transfers of genomic DNA using an adjacent single-copy fragment as probe (6)(7)(8).…”

mentioning

confidence: 99%

Propagation of some human DNA sequences in bacteriophage lambda vectors requires mutant Escherichia coli hosts.

Wyman¹,

Wolfe²,

Botstein³

1985

Proc. Natl. Acad. Sci. U.S.A.

108

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Reliable phenotyping of esterase D by low voltage isoelectric focusing: Evidence for the new variant ESD Yamaguchi

Yuasa¹,

Tamaki²,

Suenaga

et al. 1985

Electrophoresis

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Low voltage isoelectric focusing (IEF) for the classification of esterase D (ESD) has been developed. This new phenotyping method is based on non-equilibrium IEF. The obtained patterns might be isotachophoretic because the ESD bands were concentrated sharply and the values of their isoelectric point (p4 were somewhat higher than those found by equilibrium IEF. The method proved to be fast, easy and reliable for the differentiation of the six phenotypes determined by three codominant alleles, ESD* 1, ESD*2 and ESD*7. The distribution of ESD phenotypes in Western Japan has been investigated and a new rare variant, ESD Yamaguchi (ESD Y), characterized by a slightly cathodal mobility to ESD 2, could be identified. The frequencies for the three alleles ESD* 1, ESD*2 and ESD*7 were 0.600,0.389 and 0.0 1 1, respectively.

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No evidence for linkage between an insulin‐dependent diabetes mellitus‐susceptibility locus and immunoglobulin loci KM or GM

Goldstein

et al. 1984

Genetic Epidemiology

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We studied 52 families having more than one member affected with insulin-dependent diabetes mellitus (IDDM) for linkage of an IDDM-susceptibility locus to the immunoglobulin loci KM and GM. Linkage was analyzed by the LOD score method using single-locus recessive and dominant models of IDDM inheritance, with penetrances of the disease-susceptible genotypes being dependent on age and reaching a maximum of 20% by age 40. We found no evidence for linkage of IDDM to KM or GM. Close linkage (recombination fraction less than 5%) was rejected for KM under the recessive model and for GM under both models. These results suggest either that there are no IDDM-susceptibility loci closely linked to KM or GM or that use of a single-locus model of IDDM (which ignores the effects of the susceptibility locus in the HLA region) is inadequate for their detection.

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Report of the committee on the genetic constitution of chromosomes 13 to 22

Cited by 11 publications

References 6 publications

Propagation of some human DNA sequences in bacteriophage lambda vectors requires mutant Escherichia coli hosts.

Propagation of some human DNA sequences in bacteriophage lambda vectors requires mutant Escherichia coli hosts.

Reliable phenotyping of esterase D by low voltage isoelectric focusing: Evidence for the new variant ESD Yamaguchi

No evidence for linkage between an insulin‐dependent diabetes mellitus‐susceptibility locus and immunoglobulin loci KM or GM

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