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Characterization of rice α-amylase isozymes expressed by Saccharomyces cerevisiae (1995)

Terashima, M. ; Katoh, S. ; Thomas, B. R. ; [et al.]

Springer

Applied microbiology and biotechnology 43 (1995), S. 1050-1055

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ISSN: 1432-0614

Source: Springer Online Journal Archives 1860-2000

Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology

Notes: Abstract Two rice α-amylase isozymes, AmylA and Amy3D, were produced by secretion from genetically engineered strains of Saccharomyces cerevisiae. They have distinct differences in enzymatic characteristics that can be related to the physiology of the germinating rice seed. The rice isozymes were purified with immunoaffinity chromatography. The pH optima for amy3D (pH optimum 5.5) and Amy1A (pH optimum 4.2) correlate with the pH of the endosperm tissue at the times in rice seedling development when these isozymes are produced. Amy3D showed 10–14 times higher reactivity to oligosaccharides than Amy1A. Amy1A, on the other hand, showed higher reactivity to soluble starch and starch granules than Amy3D. These results suggest that the isozyme Amy3D, which is expressed at an early stage of germination, produces sugars from soluble starch during the early stage of seed germination and that the isozyme Amy1A works to initiate hydrolysis of the starch granules.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00166924

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Identification of the molecular basis for phosphorylase hypersensitivity in cultured diabetic cardiomyocytes (1995)

Buczek-Thomas, Jo Ann ; Miller, Thomas B.

Springer

Molecular and cellular biochemistry 145 (1995), S. 131-139

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ISSN: 1573-4919

Keywords: glycogen phosphorylase ; alloxan-diabetes ; cardiomyocytes ; cGMP ; phosphodiesterase

Source: Springer Online Journal Archives 1860-2000

Topics: Biology , Chemistry and Pharmacology , Medicine

Notes: Abstract The focus of this study was to identify the molecular basis for the hypersensitive response of glycogen phosphorylase activation to epinephrine stimulation in alloxan diabetic-derived cardiomyocytes. Cyclic AMP levels were found not to be significantly different between normal and diabetic-derived cells while cGMP concentrations were found consistently to be significantly lower in diabetic-derived cells than in normal cells. Treatment with cyclic GMP analogues did not affect phosphorylase activation by epinephrine in normal cardiomyocytes whereas, IBMX, a nonselective phosphodiesterase inhibitor, had a significant effect on basal and agonist-stimulated phosphorylase activity in both normal and diabetic-derived cardiomyocytes. Differences in the time course for the rate of decay of phosphorylasea from agonist-stimulated to basal levels were observed between normal and diabetic cells. After 3 h in primary culture, phosphorylasea activity returned to basal levels more quickly in normal than in diabetic-derived cells while after 24 h in culture, the time for phosphorylasea decay was not significantly different between normal and diabetic myocytes and was longer than the 3 h response. After 3 h in primary culture, no significant difference in phosphorylase kinase activity was observed between normal and diabetic-derived cells exposed to epinephrine whereas, after 24 h in culture, phosphorylase kinase activity was significantly decreased in diabetic cells under basal and agonist-stimulated conditions. These data collectively suggest that the hypersensitive response of glycogen phosphorylase to epinephrine stimulation in diabetic-derived cardiomyocytes is not due to a defect present at the level of phosphorylase kinase but may, in part, result from an alteration in cardiac phosphodiesterase activity resulting from diminished intracellular cyclic GMP concentrations.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00935485

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Genetic distribution of Bare–1-like retrotransposable elements in the barley genome revealed by sequence-specific amplification polymorphisms (S-SAP) (1997)

Waugh, R. ; McLean, K. ; Flavell, A. J. ; [et al.]

Springer

Molecular genetics and genomics 253 (1997), S. 687-694

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ISSN: 1617-4623

Keywords: Key words Bare-1 ; Retrotransposons ; Barely ; Linkage analysis ; Sequence-Specific Amplification Polymorphisms (S-SAPs)

Source: Springer Online Journal Archives 1860-2000

Topics: Biology

Notes: Abstract Retrotransposons are present in high copy number in many plant genomes. They show a considerable degree of sequence heterogeneity and insertional polymorphism, both within and between species. We describe here a polymerase chain reaction (PCR)-based method which exploits this polymorphism for the generation of molecular markers in barley. The method produces amplified fragments containing a Bare–1-like retrotransposon long terminal repeat (LTR) sequence at one end and a flanking host restriction site at the other. The level of polymorphism is higher than that revealed by amplified fragment length polymorphism (AFLP) in barley. Segregation data for 55 fragments, which were polymorphic in a doubled haploid barley population, were analysed alongside an existing framework of some 400 other markers. The markers showed a widespread distribution over the seven linkage groups, which is consistent with the distribution of the Bare–1 class of retrotransposons in the barley genome based on in situ hybridisation data. The potential applicability of this method to the mapping of other multicopy sequences in plants is discussed.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/s004380050372

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Homology of AFLP products in three mapping populations of barley (1997)

Waugh, R. ; Bonar, N. ; Baird, E. ; [et al.]

Springer

Molecular genetics and genomics 255 (1997), S. 311-321

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ISSN: 1617-4623

Keywords: Key words AFLP ; Barley ; Product homology ; Linkage maps

Source: Springer Online Journal Archives 1860-2000

Topics: Biology

Notes: Abstract Segregation of 850 polymorphic AFLP (amplified fragment length polymorphism) fragments was followed in three different doubled haploid (DH) barley populations, Dicktoo × Morex (DM), Igri × Franka (IF) and Blenheim × E224/3 (BE), which had previously been used to construct linkage maps using other molecular markers. The final maps consisted of 310, 655 and 474 markers, of which 234, 194 and 376, respectively, were AFLPs. A comparison of profiles from the parental lines identified 51 similar-sized AFLPs segregating in both DM and IF populations, 20 in the DM and BE populations and 18 in the IF and BE populations. Eight segregated in all three. Analysis of the complete datasets for each of the populations using Joinmap V.2. indicated that in general terms each of the AFLPs which were polymorphic in more than one population mapped to the same genetic locus. The number of co-dominant markers segregating in a single population ranged from 6% for DM to 12.6% for IF. These results are discussed in the context of using AFLP in genetic linkage and diversity studies.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/s004380050502

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Human cDNA clones that modify radiomimetic sensitivity of ataxia-telangiectasia (group A) cells (1995)

Ziv, Yael ; Bar-Shira, Anat ; Jorgensen, Timothy J. ; [et al.]

Springer

Somatic cell and molecular genetics 21 (1995), S. 99-111

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ISSN: 1572-9931

Source: Springer Online Journal Archives 1860-2000

Topics: Biology , Medicine

Notes: Abstract Genes responsible for genetic diseases with increased sensitivity to DNA-damaging agents can be identified using complementation cloning. This strategy is based on in vitro complementation of the cellular sensitivity by gene transfer. Ataxia-telangiectasia (A-T) is a multisystem autosomal recessive disorder involving cellular sensitivity to ionizing radiation and radiomimetic drugs. A-T is genetically heterogeneous, with four complementation groups. We attempted to identify cDNA clones that modify the radiomimetic sensitivity of A-T cells assigned to complementation group [A-T(A)]. The cells were transfected with human cDNA libraries cloned in episomal vectors, and various protocols of radiomimetic selection were applied. Thirteen cDNAs rescued from survivor cells were found to confer various degrees of radiomimetic resistance to A-T(A) cells upon repeated introduction, and one of them also partially influenced another feature of the A-T phenotype, radioresistant DNA synthesis. None of the clones mapped to the A-T locus on chromosome 11q22-23. Nine of the clones were derived from known genes, some of which are involved in cellular stress responses. We concluded that a number of different genes, not necessarily associated with A-T, can influence the response of A-T cells to radiomimetic drugs, and hence the complementation cloning approach may be less applicable to A-T than to other diseases involving abnormal processing of DNA damage.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF02255785

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