Genetic mapping of 14 short tandem repeat polymorphisms on human chromosome 22

We have constructed a linkage map of 14 short tandem repeat polymorphisms (11 with heterozygosity > 70%) on the long arm of human chromosome 22 using 23 non-CEPH pedigrees. Twelve of the markers could be positioned uniquely with a likelihood of at least 1,000:1, and distributed at an average distance of 6.62 cM (range 1.5–16.1 cM). The sex-combined map covers a total of 79.6 cM, the female map 93.2 cM and the male map 64.6 cM. Based on comparisons between physical maps and other genetic maps, we estimate that our map covers 70%–80% of the chromosome. The map integrates markers from previous genetic maps and uniquely positions one marker (D22S307). Data from physical mapping on the location of four genetic markers correlates well with our linkage map, and provides information on an additional marker (D22S315). This map will facilitate high resolution mapping of additional polymorphic loci and disease genes on chromosome 22, and act as a reference for building and verifying physical maps.     

Molecular cytogenetic analysis of a duplication Xp in a male: further delineation of a possible sex influencing region on the X chromosome

We describe a male infant with severe mental retardation and autism with a duplication of the short arm of the X chromosome. Chromosome painting confirmed the origin of this X duplication. Molecular cytogenetic analysis with fluorescence in situ hybridization (FISH) identified one copy of the zinc finger protein on the X chromosome (ZFX) and two copies of the steroid sulfatase gene (STS), further delineating the breakpoints. Based on cytogenetic and molecular comparisons of cases from the literature of sex-reversal in dup(X),Y patients and our patient, we suggest that a possible secondary sexinfluencing gene involved in the regulation of sex determination or testis morphogenesis is present at the distal Xp21.1 to p21.2 region.     

The expression of cecropin peptide in transgenic tobacco does not confer resistance to Pseudomonas syringae pv tabaci

Summary We used in vitro growth inhibition assays to demonstrate that synthetic cecropin protein has potent activity against a range of plant pathogenic bacteria. We then prepared transgenic tobacco plants which express cecropin mRNA and protein. We have used Pseudomonas syringae pv tabaci infection of these transgenic tobacco as a model system to evaluate whether the plants which express cecropin protein also have increased tolerance to infection. We found no dramatic difference in disease response between plants which are expressing cecropin protein and control plants which were derived from the transformation with a binary vector which did not carry the gene encoding cecropin protein.     

The effect of age on the recognition thresholds of three sweeteners: sucrose,saccharin and aspartame

It iS believed that people's sensitivity to taste declines with age but the evidence is inconclusive. This study was designed to test the hypothesis that taste recognition thresholds (TRTs) for sweetness are higher in older than in younger individuals, using groups of 16 younger subjects (18–30) and 16 older subjects (60–85). Three test substances were used: sucrose, aspartame and saccharin. A questionnaire recorded variables which might have affected TRTs, but data failed to show any trend that might have biased the principle variate-age. There was a significant alteration with age of recognition thresholds, at least for sucrose and saccharin. The differences between the groups for the three sweeteners were due to the fact that all the very sensitive subjects were young. None of the older subjects had particularly poor discrimination: all but one had TRTs within the range of younger subjects. Although there are age-related taste changes, they are much less dramatic than commonly occurs with other senses, such as sight and hearing. The findings of this study have implications for institutional catering and the dietary management of older people using non-sugar sweeteners.     

Development of a new transformant selection system for Penicillium chrysogenum: isolation and characterization of the P. chrysogenum acetyl-coenzyme A synthetase gene (facA) and its use as a homologous selection marker

A new transformation system for the filamentous fungus Penicillium chrysogenum is described, based on the use of the homologous acetyl-coenzyme A synthetase (facA) gene as a selection marker. Acetate-non-utilizing (Fac^–) strains of P. chrysogenum were obtained by positive selection for spontaneous resistance to fluoroacetate. Among these fac mutants putative facA strains were selected for a loss of acetyl-coenzyme A (CoA) synthetase activity. The facA gene, coding for the enzyme acetyl-CoA synthetase, was isolated from a P. chrysogenum genomic library using synthetic oligonucleotides derived from conserved regions from the corresponding genes of Aspergillus nidulans and Neurospora crassa. Vector pPC2-3, comprising a genomic 6.5 kb PstI fragment, was able to complement P. chrysogenum facA strains with frequencies up to 27 transformants·g^–1 DNA. Direct selection of transformants was accomplished using acetate and low amounts (0.001%) of glucose as carbon sources. About 50% of the transformants arose by integration of pPC2-3 DNA at the homologous facA locus and 50% by integration elsewhere in the genome. Determination of the nucleotide sequence of part of the cloned fragment showed the presence of an open reading frame of 2007 nucleotides, interrupted by five putative introns. Comparison of the nucleotide and the amino acid sequence of the facA gene of P. chrysogenum with the facA gene of A. nidulans reveals similarities of 80% and 89%, respectively. The putative introns present in the P. chrysogenum facA gene appear at identical positions as those in the A. nidulans facA gene, but show no significant sequence similarity. Correspondence to: R. F. M.van Gorcom     

Iron uptake byBifidobacterium thermophilum protoplasts

Protoplasts ofBifidobacterium thermophilum were prepared by a combination of lysozyme and protease digestion, and ferrous iron uptake studies were carried out. Little, if any, iron was internalized by the protoplasts, although large amounts of iron were bound to the protoplast surface. This binding was much greater than that of intact cells, which prefer to internalize iron by an energy-dependent process. It was also found that the binding of iron by protoplasts of cells grown in an iron-deficient medium was much more extensive than that of cells grown in an iron-sufficient medium. Soluble and particulate fractions of protoplasts were prepared by grinding them in a glass homogenizer, and the particulate fraction was also subjected to iron binding studies. The amount of iron bound was the same as that in intact protoplasts, indicating that the particulate fraction membrane fragments bound iron on their outer surface only. Nevertheless, when iron-preloaded cells were protoplasted and their surface cleared of iron, their particulate fraction contained considerable amounts of iron, indicating that the inner surface of the membranes is capable of binding iron as long as the cell is intact. The amount of iron so bound was dose-dependent on the amount of iron entering the cell. The failure of the outer and inner surface iron pools to mix was confirmed by the fact that when iron-preloaded protoplasts were incubated with additional iron, only the latter (surface-bound) was elutable with nonradioactive 2 mM FeSO₄. It is concluded that increasing bifidobacterial iron load increases the amount of iron bound to the inner surface of the membrane; the procedure, which is effective in forming bifidobacterial protoplasts, destroys their iron transport mechanism while uncovering surface iron-binding sites; and that such iron-binding sites may be of significance in the cellular iron metabolism processes.     

Structure of the intergenic spacer region from the ribosomal RNA gene family of white spruce (Picea glauca)

Five genomic clones containing ribosomal DNA repeats from the gymnosperm white spruce (Picea glauca) have been isolated and characterized by restriction enzyme analysis. No nucleotide variation or length variation was detected within the region encoding the ribosomal RNAs. Four clones which contained the intergenic spacer (IGS) region from different rDNA repeats were further characterized to reveal the sub-repeat structure within the IGS. The sub-repeats were unusually long, ranging from 540 to 990 bp but in all other respects the structure of the IGS was very similar to the organization of the IGS from wheat, Drosophila and Xenopus.     

Description and SEM Observations of Meloidogyne sasseri n. sp. (Nematoda: Meloidogynidae), Parasitizing Beachgrasses

Meloidogyne sasseri n. sp. is described and illustrated from American beachgrass (Ammophila breviliffulata) originally collected from Henlopen State Park and Fenwick Island near the Maryland state line in Delaware, United States (6). Its relationship to M. graminis, M. spartinae, and M. californiensis is discussed. Primary distinctive characters of the female perineal pattern were a high to rounded arch with shoulders, widely spaced lateral lines interrupting transverse striations, a sunken vulva and anus, and coarse broken striae around the anal area. Second-stage juvenile body length was 554 μm (470-550), stylet length 14 μm (13-14.5), tail length 93 μm (83-115), tapering to a finely rounded terminus. Male stylet length 20 μm (19-21.5), spicule length 33 μm (30-36). Scanning electron microscope observations provided additional details of perineal patterns and face views of the female, male, and J2 head. Wheat, rice, oat, Ammophila sp., Panicum sp., bermudagrass, zoysiagrass and St. Augustinegrass were tested as hosts. Distribution of the species was the coasts of Delaware and Maryland. The common name "beachgrass root-knot" is proposed for M. sasseri n. sp.     

Morphology,distribution, and preservation potential of microbial mats in the hydromagnesite-magnesite playas of the Cariboo Plateau,British Columbia,Canada

Benthic microbial mats are common in the alkaline hydromagnesite-magnesite playa lakes of Interior British Columbia. Four main zones are recognized based on mat morphology that can be related to the type and duration of wetting. From the basin margin toward the playa centre they are: (i) vegetated hummocky ground; (ii) polygonal hummocky ground; (iii) low-domal and stratiform mats, and (iv) laterally continuous and pustular mats. Mats in peripheral mudflats are commonly mineralized by hydromagnesite, mostly precipitated by capillary evaporation of shallow groundwaters. Mats forming in the ephemeral lake tend to have lower carbonate content.Although widespread, the mats are poorly preserved in the Holocene sedimentary record. Underlying sediments are commonly weakly bedded, disturbed or massive. Desiccation, dehydration, wetting-drying cycles, and grazing by invertebrates cause fragmentation of mats at the surface, facilitating erosion. Cryogranulation, interstitial mineral precipitation, vesiculation, bioturbation, compaction, and volume changes associated with diagenesis, disrupt and destroy lamination in the upper few centimetres. Most surviving organic matter is lost by early microbial degradation.     

Regional chromosomal assignment of the Kell blood group locus (KEL) to chromosome 7q33-q35 by fluorescence in situ hybridization: evidence for the polypeptide nature of antigenic variation

The gene encoding the Kell blood group polypeptide has been localized to chromosome 7q33-35 by in situ hybridization using a biotinylated 1.1-kb DNA fragment containing the 3 half of the human cDNA. This assignment is in accord with genetic localization using antigenic variation as a marker, and strongly suggests that Kell antigenic determinants are part of the polypeptide chain rather than the associated sugar molecules.