Fusion of mitochondria with protoplasts in Saccharomyces cerevisiae.

Summary Protoplasts prepared from a neutral petite haploid BO60AF-1 (a ade2 arg4 leu2 trp C ^O E ^O O ^{O O O}) were mixed with mitochondria isolated from an oligomycin resistant respiring haploid AN^ROR 12D (a his4 leu2 thr4 C ^S E ^S O _II ^{R + +}) and treated with 30% polythylene glycol and CaCl₂. When the treated protoplasts were spread and incubated on selective agar plates, oligomycin resistant respiration-sufficient colonies appeared with low frequency. All of these colonies carried the mitochondrial genotype of C ^S E ^S O _II ^{R + +} and showed the same mating type and nutritional requirements as did BO60AF-1, thus evidencing the mitochondrial transfer into protoplasts. Recombination and transmission of the mitochondrial drug resistance markers were studied in crosses involving the strains issued from mitochondria accepted protoplasts.     

Measurement of proton relaxation rates in proteins

Summary Five different types of experiment are described which make it possible to measure various relaxation rates of selected protons in crowded spectra of macromolecules such as proteins: longitudinal spin-lattice relaxation rates =1/T₁, transverse relaxation rates =1/T₂ measured under conditions of free precession, transverse relaxation rates ¹ _LOCK=1/T₁ measured under conditions of spin-locking, and transverse relaxation rates ^DQC=1/T₂ ^DQC and ^ZQC=1/T₂ ^ZQC of double- and zero-quantum coherences. The surprisingly large discrepancy between the transverse rates ^t and ^t is discussed in detail. To separate overlapping proton signals, the experimental schemes involve one or several magnetization transfer steps, using a doubly selective homonuclear Hartmann-Hahn method. Numerous variants of the basic ideas can be conceived, depending on the extent of signal overlap and on the topology of the networks of scalar couplings. Applications are shown to H and H of Tyr²³, to H, H and H of Cys³⁰, and to H and H of Ala²⁴ in bovine pancreatic trypsin inhibitor (BPTI).     

Die Aktivit?tsperiodik bei V?geln mit und ohne dunklem Schlafkasten

Zusammenfassung In 2 Versuchsserien wurden Kohlmeisen(Parus major) und japanische Möwchen(Lonchura striata var.domestica) einzeln und schallisoliert gehalten. In der ersten Versuchsserie, in der alle Vögel einen dunklen Schlafkasten hatten, wurde der Einfluß der Beleuchtungsstärke auf die Periode () der Hüpfaktivität und auf das Verhältnis von Aktivitätszeit zu Ruhezeit ( : -Verhältnis) untersucht. Sowhol Kohlmeisen als auch japanische Möwchen folgen der Regel, daß mit wachsender Beleuchtungsstärke die Periode kürzer und das : -Verhältnis größer wird.In der 2. Serie wurde der Einfluß Ruhe im dunklen Schlafkasten auf die Periodenlänge und auf das : -Verhältnis untersucht. Es wurden die Messungen aus Bedingung 1 (der Vogel hat einen dunklen Schlafkasten zur Verfügung) mit den Messungen aus Bedingung 2 (der Vogel hat keinen oder einen hellen Schlafkasten zur Verfügung) verglichen. Das Ergebnis bei Kohlmeisen entspricht den Befunden bei konstantem Licht verschiedener Intensität. Unter Bedingung 1 ist länger und das : -Verhältnis kleiner als in Bedingung 2. Das Ausmaß der Änderung von nach Fortnahme des dunklen Kastens ist unabhängig von der Periodenlänge in Bedingung 1. Das Ausmaß der änderung von : ist unabhängig von a : in Bedingung 1, jedoch schwach negativ korreliert mit der Periodenlänge in Bedingung 1.Bei japanischen Möwchen entsprechen die Ergebnisse dieser Versuchsserie nicht der Regel für tagaktive Vögel. Mit Benützen des dunklen Schlafkastens ist kürzer als ohne den Schlafkasten. Ohne den Schlafkasten ist etwa 24 Std. Das : -Verhältnis ist in Bedingung 1 unter bestimmten Voraussetzungen kleiner als in Bedingung 2. Das Ausmaß der Änderung von nach Fortnahme des Kastens ist mit der dazugehörigen Periode in Bedingung 1 hochsignifikant korreliert (Regressionskoeffizient b=-1.01, Korrelationskoeffizient r=0.89). Ebenfalls ist das Ausmaß der Änderung von : nach Fortnahme des Kastens mit : aus Bedingung 1 korreliert; es scheint, als würde ein bevorzugtes : -Verhältnis von etwa 2.0 eingeregelt.Die Ergebnisse werden im Hinblick auf 4 Punkte diskutiert: 1) Das circadiane System arbeitet innerhalb eines engen Bereiches von - und : -Werten optimal. 2) Der Optimalbereich wird bevorzugt unter ungünstigen Bedingungen angestrebt. 3) Der Entzug des dunklen Schlafkastens belastet japanische Möwchen mehr als Kohlmeisen. 4) Bei japanischen Möwchen wird in Bedingung 1 durch fortplfanzungsphysiologischen Einfluß verkürzt.

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Circadian activity rhythms of birds with and without a dark nest box

Summary Perch-hopping activity of Great tits(Parus major) and Bengales finches(Lonchura striata domestica), housed individually in soundproof boxes, was studied in two series of experiments. In the first series all birds had access to a dark nest box, in which they retired during their subjective night. In this experiment the effect of light intensity on the freerunning circadian activity rhythm was investigated. Both Great tits and Bengalese finches obey the circadian rule by responding to an increase in light intensity with shortening the circadian period () and with an increase of the ratio of activity time and rest time ( : ).In the second series of experiments the influence of sleeping in the dark nest box on both circadian period and : -ratio was studied. The results of two experimental conditions — without and with access to a dark nest box — were compared. In the Great tits, the results are in agreement with the effect of light intensity: when a dark nestbox is available, is longer and the : -ratio is smaller than in the absence of a nest box. The magnitude of the change in free-running period after removal of the nest box is independent of the original value of ; the amount of change : -ratio is likewise independent of the original : -ratio, but is weakly correlated to the original .InLonchura striata var. domestica, removal of the dark nest box leads to a lenghtening of the free-running period to about 24 hours; the : -ratio is smaller in the presence of a dark nestbox, if certain other conditions are fulfilled. The magnitude of the change in after removal of the nest box is highly correlated to the original free-running period (r=-0.89) in such a way that, without nest box, the period approaches a value of 24 hours. Also, the amount of change in : -ratio due to nest box removal is negatively correlated to the original : -ratio. A probably preferred : -ratio of 2.0 is adopted.These results are discussed in the view of 4 points: 1) The circadian system operates at its optimum within a narrow range of - and : -values. 2) This optimal range is especially adopted when conditions become adverse. 3) Removal of the dark nest box results in a more stressful situation for Bengalese finches than for Great tits. 4) In the Bengalese finches, is shortened in the presence of a nest box due to effects on reproductive physiology.

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Herrn Prof. Dr. JürgenAschoff zum 60. Geburtstag gewidemt.     

Isolation of pea nodule bacteroids utilizing silica sol (Percoll) density gradient centrifugation

Isolation of bacteroids from effective (Fix⁺) and ineffective (Fix^–) pea nodules, inoculated withRhizobium leguminosarum K, were performed by a density gradient centrifugation method using silica sol (Percoll). Only one zone (=1.064–1.072; n-zone) was recognized in the Fix⁺ nodule which contained typical Y-shaped bacteroids while two zones (n-zone and =1.125–1.145; n'-zone) were obtained from the Fix^– nodule. The cells in the n'-zone, which are long rods differed morphologically from free-living cells at any growth phase (=1.108–1.125; f-zone and =1.074–1.078; f'-zone), and differed from Y-shaped bacteroids by cell density. The esterase isozyme pattern of bacteroids in the n-and n'-zones also showed clear differences from that of f-and f'-zone of free-living cells.     

Ganglioside-cholera toxin interactions: a binding and lipid monolayer study

Summary On t.l.c. plates ¹²⁵I-cholera toxin binds to a disialoganglioside tentatively identified as GD_lb with about 10 times less capacity than to ganglioside GM₁. Binding of labeled toxin to both gangliosides was abolished in presence of excess amounts of unlabeled B subunit. Ganglioside extracts from human or pig intestinal mucosa showed toxin binding to gangliosides GM₁ and GD_1b. In ganglioside-containing lipid monolayers the penetration of the toxin was independent of the ganglioside binding capacity.Abbreviations GM₂ Gal-NAc14Gal(3-2NeuAc)14G1c1Cer - GM₁ Gal3Ga1-NAc14Gal(32NeuAc)14G1c11Cer - GD_1a NeuAc23Ga113Gal-NAc14Gal(32NeuAc)14G1c11Cer - GD1b Gall3Gal-NAcl4Gal(32NeuAc82NeuAc)14Glc11Cer - GT_1b NeuAc23Ga113Ga1-NAcal4Gal(3-2NeuAc82NeuAc)14G1c11Cer - dpPC 1,2-hexadecanoyl-sn-glycero-3-phosphocholine - dpPE 1,2-hexadecanoyl-sn-glycero-3-phosphoethanolamine     

Alternative pathway activities in aged potato tuber slices measured by three methods

To find a simple and reliable oxygen electrode-based method to estimate the values of alternative pathway activity (V _alt) and its contribution to total respiration V _alt/V _t) in aged potato (Solanum tuberosum L.) tuber slices, we compared conventional hydroxamate-inhibiting method, improved hydroxamate-inhibiting method with 2,6-dichlorophenol indophenol (DCPIP), and the oxygen isotope discrimination (OID) method. The values of V _alt and V _alt/V _t obtained with an improved hydroxamate-inhibiting method with DCPIP in 12-h- and 24-h-aged slices were about twice higher than those with the conventional hydroxamate-inhibiting method. Only a relatively small difference in the values of V _alt and V _alt/V _t obtained by the OID method and the improved hydroxamate-inhibiting method with DCPIP in 12-h and 24-h-aged slices was observed. These results indicated that the improved hydroxamate-inhibiting method with DCPIP could be considered as a new, simple, and reliable technique for the noninvasive assay of the AP activity.From Fiziologiya Rastenii, Vol. 52, No. 2, 2005, pp. 311–315.Original English Text Copyright © 2005 by Hou, Zhou, Kong, Liang, Zhang.This article was submitted by the authors in English.This revised version was published online in April 2005 with a corrected cover date.     

Bacteriophage phi 1 as a gene-cloning vector in Bacillus subtilis

Summary We attempted to use Bacillus subtilis phage 1 as a gene-cloning vector since the 1 genome was found to have few cleavage sites upon digestion with several kinds of restriction endonucleases. A 1 stock supplied by J. Ito (University of Arizona, Tucson, USA) consisted of two phages, 1E1 and 1E2, having one and two EcoRI-cleavage sites in their genomes respectively. From the latter isolate a deletion mutant 1E21 was induced to increase the size range of DNA segments to be cloned. It was demonstrated, by in vitro recombination experiments with phage 11 DNA, that 1E21 can be used for cloning EcoRI fragments of various sizes. We analyzed the DNAs of ten 1 clones isolated from independent transfectants and found that six of them carried 11 DNA fragments inserted at either of the two EcoRI-cleavage sites. Some of the hybrid phage DNAs were found to be cleaved with BamHI and HaeIII endonucleases at the 11 DNA portion, whereas the parental 1E21 DNA was insensitive to any of these enzymes. These hybrid phages would therefore be useful vectors for cloning foreign DNA fragments generated by cleavage with BamHI or HaeIII endonucleases.     

Mutants in yeast affecting ethidium bromide induced rho- formation and their effects on transmission and recombination of mitochondrial genes.

Summary A series of mutants called ebi, less inducible by ethidium bromide than the parental strain for the ^+– mutation have been isolated after E.M.S. mutagenesis. Some of the ebi mutants also show an important accumulation of ^– cells, in the absence of ethidium bromide. Ebi mutations are nuclearly inherited as shown by meiotic segregation. The effects of these mutants on the transmission and recombination of mitochondrial genes among the diploid progeny of crosses have been studied. Some of the ebi mutants show a non coordinated transmission of the oli1 mitochondrial marker with respect to other mitochondrial markers unexpected for homosexual crosses. This bias which is independent from will be discussed in relation to the segregation and recombination. No significant decrease of the frequency of recombinants has been detected.Abbreviations E.B. Ethidium bromide - E.M.S. Ethyl méthane sulfonate - C^S/C^R Allelic forms of the rib 1 locus conferring chloramphenicol sensitivity/resistance - E^S/E^R Allelic forms of the rib 3 locus conferring erythromycine sensitivity/resistance - O^R/O^R Allelic forms of the oli 1 locus conferring oligomycin sensitivity/resistance - P^S/P^R Allelic forms of the par 1 locus conferring paromomycine sensitivity/resistance - D^S/D^R Allelic forms of the diu 1 or diu 2 loci conferring diuron sensitivity/resistance - C^S/C^R Allelic forms of the mitochondrial locus - ⁺ grande or respiratory competent cells - ^– petite or cytoplasmic respiratory deficient cells     

Variable generation times and Darwinian fitness measures

Summary Reproductive value (RV) and net reproductive output (R _o) are frequently used fitness measures. We argue that they are only appropriate when intervals between reproductive events are fixed, as they are dimensionless generation-to-generation scalings with units offspring per parent. A fitness measure should account for two different effects of a decrease in generation time: (1) increased survival due to shorter exposure to mortality agents and (2) increased frequency of reproduction.R _o andRV deal with the first of these two effects, while a measure with a physical dimensionper time [T^–1] is needed to account for the second. The Malthusian growth parameter,r, meets this requirement and in situations where time to reproduction is variable, we propose, the instantaneous rate of spread of descendants (from an individual) be used instead ofR _o. As an alternative toRV, we suggest using the instantaneous difference = –r, wherer is the population rate of increase. WhileRV andR _o are dimensionless ratios, , and areper time rates which are appropriate in accounting for alterations in generation time.     

Enzymic synthesis of the trisaccharide core region of the carbohydrate chain ofN-glycoprotein

Transmannosylation from mannotriose (Man1-4Man1-4Man) to the 4-position at the nonreducing end N-acetylglucosaminyl residue ofN,N-diacetylchitobiose was regioselectively induced through the use of -d-mannanase fromAspergillus niger. The enzyme formed the trisaccharide Man1-4GlcNAc1-4GlcNAc (3.7% of the enzyme-catalysed net decrease ofN,N-diacetylchitobiose) from mannotriose as a donor andN,N-diacetylchitobiose as an acceptor. Mannobiose (Man1-4Man) was also shown to be useful as a donor substrate for the desired trisaccharide synthesis.Abbreviations Man d-mannose - (M _n) (n=1–5) -linkedn-mer of mannose - GlcNAc₂ 2-acetamido-2-deoxy--d-glucopyranosyl-(1–4)-2-acetamido-2-deoxy-d-glucose     

Modelling of predator–prey trophic interactions. Part I: two trophic levels

A class of lumped parameter models to describe the local dynamics in a controlled environment of a two-trophic chain is considered. The class is characterized by a trophic function (functional response of predator to the abundance of prey) depending on the ratio of prey biomass x and a linear function of predator biomass y: f(qx/[(1-)k+y]), where q is the efficiency of the predation process, k is a reference biomass, and (01) specifies the predation model. The trophic function is defined only by some properties determining its shape. A stability analysis of the models has been performed by taking the parameters q and as bifurcation parameters: the regions in the (,q) plane of existence and stability of nonnegative equilibrium states and limit cycles are determined. This analysis shows that the behaviour of the models is qualitatively similar for 0<1 (in particular the null state is always a saddle point), while the value =1 gives rise to some kind of structural instability of the system (in particular the null state becomes an attractor for sufficiently high predation efficiency).     

Hydrogen isotope discrimination in higher plants: Correlations with photosynthetic pathway and environment

Summary The ratio of deuterium to hydrogen (expressed as D) in hydrogen released as water during the combustion of dried plant material was examined. The D value (metabolic hydrogen) determined on plant materials grown under controlled conditions is correlated with pathways of photosynthetic carbon metabolism. C₃ plants show mean D values of-132 for shoots and -117 for roots; C₄ plants show mean D values of -91 for shoots and-77 for roots and CAM plants a D value of-75 for roots and shoots. The difference between the D value of shoot material from C₃ and C₄ plants was confirmed in species growing under a range of glasshouse conditions. This difference in D value between C₃ and C₄ species does not appear to be due to differences in the D value (tissue water) in the plants as a result of physical fractionation of hydrogen isotopes during transpiration. In C₃ and C₄ plants the hydrogen isotope discrimination is in the same direction as the carbon isotope discrimination and factors contributing to the difference in D values are discussed. In CAM plants grown in the laboratory or collected from the field D values range from-75 to +50 and are correlated with ¹³C values. When deprived of water, the D value (metabolic hydrogen) in both soluble and insoluble material in leaves of Kalanchoe daigremontiana Hamet et Perr., becomes less negative. These changes may reflect the deuterium enrichment of tissue water during transpiration, or in field conditions, may reflect the different D value of available water in areas of increasing aridity. Whatever the origin of the variable D value in CAM plants, this parameter may be a useful index of the water relations of these plants under natural conditions.     

Average phase difference theory and 1∶1 phase entrainment in interlimb coordination

The dynamics of coupled biological oscillators can be modeled by averaging the effects of coupling over each oscillatory cycle so that the coupling depends on the phase difference between the two oscillators and not on their specific states. Average phase difference theory claims that mode locking phenomena can be predicted by the average effects of the coupling influences. As a starting point for both empirical and theoretical investigations, Rand et al. (1988) have proposed d/dt= — K sin ), with phase-locked solutions =arcsin( /K), where is the difference between the uncoupled frequencies and K is the coupling strength. Phase-locking was evaluated in three experiments using an interlimb coordination paradigm in which a person oscillates hand-held pendulums. was controlled through length differences in the left and right pendulums. The coupled frequency _c was varied by a metronome, and scaled to the eigenfrequency _v of the coupled system K was assumed to vary inversely with _c. The results indicate that: (1) and K contribute multiplicatively to (2) =0 or = regardless of K when =0; (3) 0 or regardless of when K is large (relative to ); (4) results (1) to (3) hold identically for both in phase and antiphase coordination. The results also indicate that the relevant frequency is _c/_v rather than _c. Discussion high-lighted the significance of confirming =arcsin(/K) for more general treatments of phase-locking, such as circle map dynamics, and for the 11 phase-entrainment which characterizes biological movement systems.     

Respiratory activity and growth kinetics of Candida yeasts related to carbon sources and available energy

Summary Three yeasts of the genus Candida (Candida intermedia, candida lipolytica and Candida tropicalis) were cultivated batchwise on three different carbon sources: glucose, acetate, and hexadecane. Growth curves, oxygen uptake rates, CO₂ evolution rates and the amount of oxygen required for biomass production were determined. The data were compared and discussed from the point of maximum specific growth rate, maximum oxygen uptake rate, carbon conversion into CO₂ and biomass, consumption of oxygen and available energy for cell synthesis. The results indicated a relationship between _m m, Y_s, Y_O, and for different carbon sources. Y_O and were in the same order of magnitude for acetate (0.58 and 0.38 respectively) and hexadecane (0.45 and 0.40 respectively). These values were remarkably lower than those for glucose (1.26 and 0.54 respectively).Symbols av e^– Available electrons per mol of substrate (dimensionless) - E_av Energy available per mol of substrate (dimensionless) - C_d Dissimilated carbon (%) - m Maximum specific rate of oxygen uptake (mMO₂ h^–1 g^–1) - RQ CO₂ evolved per O₂ consumed - mol. wt. Molecular weight - Y_ATP Biomass mass yield based on mol of ATP generated (g) - Biomass mass yield based on available energy (g) - Y_M Biomass mass yield based on mol of organic substrate (g) - Y_O Biomass mass yield based on oxygen consumed (gg^–1) - 1/Y_O Oxygen consumed for one gram of biomass produced (gg^–1) - Y_s Biomass mass yield based on organic substrate (dimensionless) - b Reductance degree of biomass (equiv. available electrons/g atom carbon) - s Reductance degree of organic substrate (equiv. available electrons/g atom carbon) - Fraction of energy in organic substrate which is converted to biomass - b Weight fraction carbon in biomass (dimensionless) - s Weight fraction carbon in organic substrate (dimensionless) - _m Maximum specific growth rate (h^–1)     

A quantitative method for separating reaction components of CMP-sialic acid: Lactosylceramide sialyltransferase using Sep Pak C18 cartridges

A rapid procedure is described for the separation of CMP-sialic acid:lactosylceramide sialyltransferase reaction components using Sep Pak C₁₈ cartridges. The quantitative separation of the more polar nucleotide sugar, CMP-sialic acid, and its free acid from the less polar G_M3-ganglioside is simple and rapid relative to previously described methods. Recovery of G_M3 is optimized by the addition of phosphatidylcholine to the reaction mixture prior to the chromatographic step. Using rat liver Golgi membranes as a source of CMP-sialic acid: lactosylceramide sialyltransferase activity (G_M3 synthase; ST-1), the transfer of [¹⁴C] sialic acid from CMP-[¹⁴C] sialic acid to lactosylceramide can be quantified by this assay. The procedure is reliable and may be applicable to the isolation of ganglioside products in otherin vitro glycosyltransferase assays.Abbreviations G_M3 G_M3-ganglioside - II³NeuAc-LacCer NeuAc2-3Gal1-4Glc1-1Cer - G_D1a G_D1a-ganglioside, IV³NeuAc, II³NeuAc-GgOse₄Cer, NeuAc2-3Gal1-3GalNac1-4(NeuAc2-3)Gal1-4Glc1-1Cer - G_D3 G_D3-ganglioside, II³(NeuAc)₂LacCer, NeuAc2-8NeuAc2-3Gal1-4Glc1-1Cer - GgOse₄Cer asialo-G_M1 Gal1-3GalNAc1-4Gal1-4Glc1-1Cer - FucG_MI fucosyl-G_MI-ganglioside, Fuc1-2Gal1-3GalNAc1-4Gal1-4 Glc1-1Cer - ST-1 G_M3 synthase, CMP-sialic acid:lactosylceramide sialyltransferase - LacCer lactosylceramide, Gal1-4Glc1-1Cer - CMP-NeuAc cytidine 5-monophospho-N-acetylneuraminic acid - PC phosphatidylcholine - PMSF phenylmethylsulfonyl fluoride     

Off-resonance R1rho relaxation outside of the fast exchange limit: an experimental study of a cavity mutant of T4 lysozyme

An ¹⁵N off-resonance R ₁ spin relaxation study of an L99A point mutant of T4 lysozyme is presented. Previous CPMG-based relaxation dispersion studies of exchange in this protein have established that the molecule interconverts between a populated ground state and an excited state (3.4%) with an exchange rate constant of 1450 s^–1 at 25°C. It is shown that for the majority of residues in this protein the offset dependence of the R ₁ relaxation rates cannot be well fit using models which are only valid in the fast exchange regime. In contrast, a recently derived expression by Trott and Palmer (J. Magn. Reson., 154, 157–160, 2002) which is valid over a wider window of exchange than other relations, is shown to fit the data well. Values of (signed) chemical shift differences between exchanging sites have been extracted and are in reasonable agreement with shift differences measured using CPMG methods. A set of simulations is presented which help establish the exchange regimes that are best suited to analysis by off-resonance R ₁ techniques.     

Changes of fluorescence lifetime and rotational correlation time of bovine serum albumin labeled with 1-dimethylaminonaphthalene-5-sulfonyl chloride in guanidine and thermal denaturations

The nanosecond fluorescence depolarization method was applied to measure the fluorescence lifetime () and the rotational correlation time () of bovine serum albumin (BSA) labeled with 1-dimethylaminonaphthalene-5-sulfonyl chloride (dansyl-Cl). Changes of and of dansyl BSA in the guanidine denaturation and in the thermal denaturation were examined. In parallel, the secondary structural change of dansyl BSA was followed by circular dichroism measurements. The magnitude of was almost unchanged between 1 and 2 M guanidine, where the secondary structure of the protein was predominantly disrupted; whereas that of began to increase before the disruption of secondary structure in the guanidine denaturation. In the thermal denaturation, in contrast, changes of both and occurred in a temperature range where the secondary structure was predominantly disrupted. The volume of equivalent sphere (V _e ) and the axial ratio () for the BSA were 3.6–3.8×10^–19 cm³ and 3.6 at 2M guanidine as against 2.1×10^–19 cm³ and 2.2 in the absence of guanidine (25°C), respectively. The magnitudes ofV _e and were 4.9×10^–19 cm³ and 4.5 at 65°C, respectively. Although the secondary structural change of dansyl BSA was irreversible in the thermal denaturation,V _e and were reversible.     

Detection of specific DNA sequences in yeast by colony hybridization

Summary A procedure is described for the detection of specific DNA sequences in Saccharomyces cerevisiae. This method allows a rapid screening of a large number of yeast colonies. The yeast cells of each colony, grown on nitrocellulose filters, are converted, in situ, to protoplasts by snail enzyme, and are then lysed and their DNAs are denatured and fixed on the filter. The presence of the specific DNA sequence is detected directly on the filter by hybridization with a radioactive cRNA.We have used successfully this technique to detect the presence or the absence of specific mt DNA sequences, in ⁺, ^- and ⁰ strains, and to detect the presence or the absence of the 2 m DNA sequences in different strains.     

Orientation of the porphyrin ring in artificial chlorophyll membranes

Summary A sensitive photometric method is described by which the dichroism of lipid bilayer membranes in aqueous phase can be measured. The method is applied to black films with incorporated chlorophylla andb. With chlorophylla a relatively large dichroism is found in the Soret band and a much weaker dichroism in the red band. From the experimental data, the angles _B and _R between the blue and red transition moments and the membrane can be obtained. _B and _R are then used to calculate the angle of the porphyrin ring with respect to the membrane surface. For chlorophylla and three different lipids, values of between 44 and 49° are found.     

Assortative mating and the genetic correlation

Summary The effect of assortative mating on the genetic correlation between traits X and Y is considered. Assortation on trait X changes the magnitude of the genetic correlation but not its sign. There are two situations depending on the signs of the correlation between mates () and of the random mating genetic correlation (): 1) if sign () = sign (), then >, where is the genetic correlation at equilibrium after continued assortation, and 2) if sign () = sign (), then < . However, negative assortative mating is virtually powerless to alter the magnitude of the genetic correlation. The consequences of a mixed assortation model, e.g., high milk production females mated to fast growing males and lesser productive females mated to slower growing sires, were also studied. Mixed positive assortation always increases the genetic correlation, but negative assortation decreases it. The implications of assortative mating on correlated responses to selection and on the equilibrium covariances between relatives for pairs of traits are discussed.