Allozyme variation in the stingless bee Trigona fuscobalteata (hymenoptera,apidae) from Peninsular Malaysia

• 1.1. A population of Trigona fuscobalteata from Peninsular Malaysia was analysed for genetic variation at 9 gene-enzyme systems comprising 13 loci.
• 2.2. Two gene-enzyme systems (phosphoglucomutase and isocitrate dehydrogenase) were polymorphic in the 20 colonies studied.
• 3.3. Isocitrate dehydrogenase was represented by duplicate genes.
• 4.4. The number of loci for several enzyme systems appeared to be different from that reported for the Australian stingless bees.

     

Comparative examination of the soluble muscle proteins of two species of angler-fish (Lophiidae): Lophius piscatorius (L.) and Lophius budegassa (Spinola)

• 1.1. A comparative examination of sarcoplasmic proteins of the two nominal European species of angler-fish, Lophius piscatorius and L. budegassa was carried out using isoelectric focusing techniques.
• 2.2. Two protein bands differing in isoelectric point proved diagnostic for L. budegassa (pI 4.40 and pI 5.75) while a third characterized L. piscatorius (pI 4.65).
• 3.3. These species-specific protein profiles provide a method of species discrimination independent of morphological criteria.
• 4.4. Within-species heterogeneity of banding pattern suggested the presence of polymorphic gene loci of potential use in studies of population structure.

     

Biochemical evidences for two sibling species of genus Myliobatis (chondrichthyes: myliobatidae) in South Brazil

• 1.1. Two morphotypes of Myliobatis from the demersal fishery off the Rio Grande (Brazil) were studied.
• 2.2. Thirty-two alleles were detected and resolved by 27 loci.
• 3.3. Nei's measure of genetic identity was 0.8306 and Thorpe's similarity was 0.6990. Mean heterozygosities observed were 0.1327 for the “DE” morphotype and 0.0409 for the “DL” morphotype.
• 4.4. Seven loci were fixed differently in the two taxa studied. This indicates the existence of a barrier to gene-flow between them, showing that both morphotypes belong to different species.
• 5.5. Jaccard's measure of similarity was calculated and a phenogram with the two morphotypes and M. freminvillii was constructed using isoelectric focusing of total soluble proteins. This showed a higher similarity between the two morphotypes of Myliobatis than M. freminvillii.

     

Allozyme variation in a wild african elephant (Loxodonta africana) population from the Kruger National Park,South Africa

• 1.1. Blood, liver, heart, testis, skin, eye, muscle and kidney samples were obtained from elephants (Loxodonta africana) in the Kruger National Park during a culling programme in April 1992.
• 2.2. Gene products of 25 protein coding loci in L. africana were examined by horizontal starch-gel electrophoresis.
• 3.3. Eighteen protein coding loci (72%) displayed monomorphic gel banding patterns whereas only seven (28%) displayed polymorphic gel banding patterns.
• 4.4. Average heterozygosity values for adults, youngsters and the total population are respectively 0.058, 0.024 and 0.047.
• 5.5. Relative gene diversities within and between populations are 84% and 16% respectively.
• 6.6. Two population simulation programmes were utilized to predict the duration of the current variability present in this species, based on current genetic variation and gene transfer from one generation to the next.

     

Allozyme variation in a dimeric esterase of the oriental fruit fly,Dacus dorsalis (insecta: tephritidae), from peninsular malaysia

• 1.1. Seven natural populations of Dacus dorsalis were analyzed for a dimeric esterase by means of horizontal starch-gel electrophoresis.
• 2.2. The electrophoretic phenotypes were governed by nine codominant Est-D alleles.
• 3.3. The commonest allele in all seven population samples was Est-D¹⁰⁰ which encoded an electrophoretic band with intermediate mobility.
• 4.4. The distribution of EST-D phenotypes were in accordance with Hardy-Weinberg expectations.
• 5.5. There was no geographic variation in the distribution of Est-D alleles.

     

The suitability of using cryopreservation of spermatozoa for the conservation of genetic diversity in African catfish (Clarias gariepinus)

• 1.1. The effect of cryopreservation of semen on expected Hardy-Weinberg proportions in the f₁ progeny of selected breeding pairs was evaluated.
• 2.2. Four different African catfish breeding pairs were selected, each pair displaying different heterozygous alleles at the glucose-6-phosphate isomerase-1 or 2 loci.
• 3.3. Equal volumes of ova from each female were artificially inseminated with cryopreserved or fresh semen obtained from males possessing corresponding genotypes.
• 4.4. A comparison of growth rates between f₁ groups of offspring produced from fresh and cryopreserved semen was made.
• 5.5. The G-test for goodness of fit showed no significant differences from expected Hardy-Weinberg proportions for allele frequencies obtained in the f₁ progeny.
• 6.6. The application of cryopreservation for the conservation of genetic diversity is discussed.

     

Electrophoretic identification of bird species involved in collisions with aircraft

• 1.1. The utility of biochemical genetic methods of bird identification was investigated for some common species which create a hazard for commercial aviation in Ireland.
• 2.2. Sixteen enzyme loci were assayed in eight species, using starch gel electrophoresis; three larids, three corvids and two columbids.
• 3.3. Genera were distinguishable using all but two loci.
• 4.4. Differences within genera were small, but all species except for the gulls Larus argentatus and L. marinus, could be identified using one or more loci.
• 5.5. Arising from the success of the method using fresh specimens, a protocol for the electrophoretic identification of traumatized remains of strikes is suggested.

     

Genetic variability in south african blue wildebeest (Connochaetes taurinus)

• 1.1. We used protein gel-electrophoresis to investigate genetic heterogeneity at 33 protein coding loci in a total of 46 blue wildebeest (C. taurinus) kept under different management regimes.
• 2.2. Average heterozygosity ranged from 2.14 to 4.3% and within-population differences accounted for 97.2% of total relative gene diversity.
• 3.3. Comparatively little divergence was found between animals sampled from populations with very diverse population sizes and management histories, with the largest genetic distance estimated between any two populations being only 0.0021.
• 4.4. We discuss our results with particular emphasis on the influence of management history on genetic diversity and divergence in C. taurinus.

     

An electrophoretic study of genetic variation in populations of Yarrow's spiny lizard Sceloporus jarrovi (Sauria,Iguanidae)—I. Esterae characterization and polymorphism

• 1.1. Muscle esterase variation in Sceloporus jarrovi, sampled from 25 locations in southeastern Arizona, was investigated employing acrylamide gel electrophoresis.
• 2.2. Three distinct esterase phenotypes were observed, presumably resulting from the expression of two gene loci, Est-1 and Est-2.
• 3.3. Lizards sampled from all 25 locations were found to be monomorphic with respect to esterase encoded at Est-1. Further, lizards sampled from the Santa Rita and Pinaleno Mountains were also found to be monomorphic for esterases encoded at Est-2, whereas those sampled from the Chiricahua and Huachuca Mountains proved to be polymorphic.
• 4.4. Characterization of the esterases utilizing eserine sulfate, diisopropylfluorophosphate, and sulfhydryl-group inhibitors revealed the EST-1 isozyme to be an arylesterase and the EST-2 isozymes to be carboxylesterases.

     

Malate dehydrogenase isoforms from ribbed mussel (Modiolus demissus) gill tissue

• 1.1. The malate dehydrogenase (MHD) activity from the ribbed mussel gill is polymorphic with two distinct mitochondrial forms (M1 and M2) and five forms that could be resolved from cytosolic extracts (C1 to C5) by DEAE-cellulose chromatography and starch gel electrophoresis.
• 2.2. Two of the cytosolic forms (C3 and C4) may represent interchangeable conformational states.
• 3.3. With kinetic analysis there appear to be three distinct cytosolic forms (C1, C2 and C3–C4), with C2 possibly behaving as a heterodimer.
• 4.4. The identity of C5 is uncertain.
• 5.5. The forms isolated from the mitochondria (M1 and M2) exhibited lower apparent K_ms for oxaloacetate (OAA) than the cytosolic forms.
• 6.6. For all isozymic forms, the apparent K_ms for OAA increased as the pH increased between pH 6 and 9
• 7.7. Increasing the salt concentration raised the K_m for OAA for all forms.
• 8.8. The mMDHs were more sensitive to inhibition by NaCl than the cMDHs.
• 9.9. Representative cMDH (C1) and mMDH (M2) isozymes exhibited substrate inhibition by high concentrations of OAA with the mMDH possessing lower K_is for substrate inhibition than the cMDH at each pH tested.
• 10.10. Differences and similarities in K_m app. for OAA at the different pHs and salt concentrations indicated that C1, C2 and C3–C4 and C5 were distinct forms, that M1 and M2 were distinct but very similar to each other, and that C1, C2, C3–C4 and C5 were distinct from M1 and M2.

     

A new glutamate dehydrogenase from Halobacterium halobium with different coenzyme specificity

• 1.1. Halobacterium halobium has two chromatographically distinct forms of glutamate dehydrogenase which differ in their thermolability and other properties. One glutamate dehydrogenase utilizes NAD, the other NADP as a coenzyme.
• 2.2. The NADP-specific glutamate dehydrogenase (EC 1.4.1.4) was purified 65-fold from crude extracts of H. halobium.
• 3.3. The Michaelis constants for 2-oxoglutarate (13.3 mM), ammonium (3.1 mM) and NADPH (0.077 mM) indicate that the enzyme catalyzes in vivo the formation of glutamate from ammonium and 2-oxoglutarate.
• 4.4. The amination of 2-oxoglutarate by NADP-specific glutamate dehydrogenase is optimal at the pH value of 8.0–8.5. The optimal NaCl or KCl concentration for the reaction is 1.6 M.
• 5.5. None of the several metabolites tested for a possible role in the regulation of glutamate dehydrogenase activity appeared to exert an appreciable influence on the enzyme.
• 6.6. NAD- and NADP-dependent glutamate dehydrogenases from H. halobium showed apparent molecular weights of 148,000 and 215,000 respectively.

     

Electrophoretic characterization of a hybrid between Eretmochelys imbricata and Caretta caretta (Cheloniidae)

• 1.1. An intermediate morphotype between Eretmochelys imbricata and Caretta caretta was studied in Praia do Forte, Bahia, Brazil.
• 2.2. Three enzymatic systems were successfully analyzed: SOD, lactate dehydrogenase (LDH) and esterase (EST). Isoelectric focusing of total soluble proteins of muscle and transferrin were shown.
• 3.3. Esterase exhibited nine phenotype patterns, seven in C. caretta and one in the others morphotypes. SOD phenotypes were identical in the three morphotypes. Lactate dehydrogenase and transferrins were characteristic for each species.
• 4.4. Jaccard's measure of similarity was calculated and a phenogram with the three morphotypes were constructed using isoelectric focusing of total soluble proteins.

     

Light-induced effects of several enzymes of carbohydrate metabolism in Phycomyces blakesleeanus

• 1.1. The photoregulation shown by glyceraldehyde 3-phosphate dehydrogenase and glucose 6-phosphate dehydrogenase appears to be independent of the mad gene product(s) and also independent of carotene biosynthesis regulation.
• 2.2. The photoregulation of malate dehydrogenase appeared to be dependent on the mutation of the mad and car S genes.
• 3.3. Pyruvate kinase and lactate dehydrogenase may be classified as light-independent.
• 4.4. The action of ATP and fructose 1,6-bisphosphate on the enzymes studied was generally independent of light/dark grown conditions.
• 5.5. However, the effect of fructose 1,6-bisphosphate on Phycomyces pyruvate kinase appears to be light-dependent.

     

Purification and some properties of isocitrate dehydrogenase of a blowfly Aldrichina grahami

• 1.1. NADH-dependent isocitrate dehydrogenase has been purified 110-fold from the crude extract of the flight muscle mitochondria of Aldrichina grahami.
• 2.2. The purification procedure involved Triton X-100 treatment of isolated mitochondria, column chromatography on DEAE-cellulose, Affi-gel blue, and P-cellulose.
• 3.3. The purified enzyme was homogeneous by criteria of the polyacrylamide gel electrophoresis.
• 4.4. The enzyme of the blowfly contains more acidic amino acids and less hydrophobic amino acids than that of pig heart.
• 5.5. The molecular weight was determined to be 330,000 daltons. The subunit construction differs from ghat of mammalian isocitrate dehydrogenase.

     

Distribution of beta-adrenergic binding in fractionated porcine adipocytes

• 1.1. The subcellular distribution of the porcine adipocyte beta-adrenergic receptor was studied in fractionated adipocytes.
• 2.2. The 30,000 g pellet obtained from hypotonically lysed cells contained membrane vesicles and mitochondria; it yielded approx 200–300 fmol dihydroalprenolol-bound receptors/mg protein.
• 3.3. Activity was increased to about 1000 fmol/mg protein after isolation of a plasma membrane fraction on a Percoll gradient.
• 4.4. The 5'-nucleotidase, succinate dehydrogenase and lactate dehydrogenase activities were usually enriched in compartments different from the ligand-binding activity.
• 5.5. Activity of porcine adipocyte 5'-nucleotidase, a purported plasma membrane marker enzyme, was not distributed in the same manner as the beta-adrenergic receptor.

     

Genetic variation in the nine rainbow trout (Oncorhynchus mykiss) populations in South Africa

• 1.1. Twenty-six protein coding loci in nine populations of rainbow trout, from different localities in South Africa, were analyzed by starch and polyacrylamide gel electrophoresis to estimate the amount of genetic diversity present.
• 2.2. Heterozygosity ranged between 0.0445 and 0.0745.
• 3.3. The relatively high heterozygosity levels are probably due to the historical mixing of strains, and the differences in heterozygosity indicate that the populations originate from different sources, each with different genetical characteristics.
• 4.4. Differences in phenotypic characters (i.e. growth) seem to relate to genotypic variation, whilst food conversion rate and survival performance results did not seem to relate to genotypic variation.

     

Morphometric and electrophoretic evidences for two species of the genus Liopetrolisthes (crustacea: decapoda: porcellanidae) and some aspects of their variability

• 1.1. Four colour forms of the porcellanid commensal crab Liopetrolisthes from Chilean coastal waters were compared by canonical discriminant analysis (DCA) over 10 morphological variables and by horizontal starch gel electrophoresis of 16 enzymes and total proteins with the aim of determining whether they correspond to the single species L. mitra or to different species.
• 2.2. The morphometric analysis showed the existence of two morphological groups which were principally discriminated by the dactylus and propodus dimensions of the chelipods.
• 3.3. Nine enzymatic systems were successfully tested: α-glycerophosphate dehydrogenase (α-GPDH), malic enzyme (ME), alkaline phosphatase (ALP), phosphoglucose isomerse (PGI), phosphoglucomutase (PGM), glutamate-oxalacetate transaminase (GOT), isocitrate dehydrogenase (IDH), lactae dehydrogenase (LDH), malate dehydrogenase (MDH) and total proteins (PT), which gave a total of 16 loci analyzed.
• 4.4. The biochemical analysis demonstrated that there were no electrophoretic differences within the two morphological groups discriminated by the DCA. However there was great genetic divergence among them, expressed by eight diagnostic loci and by an unbiased estimate of Nei's [Gentics, Austin, Texas 89, 583–590 (1978)] genetic distance of 0.979.
• 5.5. The magnitude of this divergence and the lack of evidence of gene flow between the two morphological groups, allows us to conclude that Liopetrolisthes shows two valid species.
• 6.6. A following study [Weber, Gayana Zool. (in press, 1991)] demonstrated that these two species are L. mitra [Dana, United States Exploring Expedition during the years 1838–1842, under the command of Charles Wilkes, U.S.N., Vol. 13, Crustacea, Part 1, p. 685 (1852)] and L. patagonicus [Cunningham, Trans. Linn. Soc. London27, 465–502 (1871)].
• 7.7. Liopetrolisthes mitra showed a polymorphism of 0.200 and an unbiased observed heterozygosity of 0.089 which are nearly values given for anomurans. Liopetrolisthes patagonicus, which lives on the overexploited sea-urchin Loxechinus albus, showed much reduced variability with a polymorphism of 0.066 and an unbiased mean observed heterozygosity of 0.008, which may be related to the fishing pressure-bottleneck hypothesis.

     

Role of glutamate dehydrogenase reaction in the control of citrate pool in yeast

• 1.1. Role of NADP-glutamate dehydrogenase in the depletion of citrate was analyzed using permeabilized yeast cells.
• 2.2. Citrate was converted to 2-oxoglutarate, which was then metabolized to glutamate by NADP-glutamate dehydrogenase in the presence of ammonium ion.
• 3.3. Formation of 2-oxoglutarate plus glutamate was in good agreement with the concentration of citrate decreased. Glutamate formation can be a good indicator of the depletion of citrate, because 70% of the citrate decreased was converted to glutamate.
• 4.4. Glycolytic activity was closely correlated with the decrease in citrate under the in situ conditions.
• 5.5. NADP-glutamate dehydrogenase increased in anaerobically grown yeast cells.
• 6.6. An effective depletion of citrate by increased synthesis of NADP-glutamate dehydrogenase can explain the lowered mechanism of citrate causing glycolytic stimulation under the anaerobic growth conditions of yeast.

     

Purification and characterization of the d-lactate dehydrogenase from Leuconostoc lactis

• 1.1. The d-lactate dehydrogenase from Leuconostoc lactis has been purified in high yield.
• 2.2.The enzyme is a dimer of subunits of M_r = 39,000 and each subunit contains a single thiol group. The N-terminal residue is methionine.
• 3.3. The amino acid composition has been determined and is typical of that of a soluble globular protein.

     

The regulation of glucose 6-phosphate dehydrogenase from Dicentrarchus labrax (bass) liver

• 1.1. Kinetic constant values of the reaction catalyzed by bass liver glucose 6-phosphate dehydrogenase show to be modified between 10 and 40°C.
• 2.2. The Arrhenius plot between 10 and 50°C shows two slopes with different activation energies.
• 3.3. These results suggest a regulation of this enzyme by environmental temperature.
• 4.4. Kinetics of ATP inhibition were examined between pH 6.2 and 7.8: patterns and K_i values obtained are affected by the pH variation.
• 5.5. NADH is an effective inhibitor of bass glucose 6-phosphate dehydrogenase but this enzyme does not show NAD-linked activity.
• 6.6. Kinetics of pyridoxal 5′-phosphate inhibition have indicated the presence of a lysine in the catalytic site for NADP⁺.