Ti质粒T区tms基因的分离及其对高等植物分化的影响

根癌农杆菌Ti质粒的T区DNA带有致瘤基因,其基因1和基因2编码生长素吲哚乙酸生物合成途径中的两个酶。以pGV 354(pBR322质粒中插有Ti质粒C 58 T区DNA的HindⅢ15—HindⅢ22大片段)重组质粒出发,我们分离了基因1和基因2,并构建了带有卡那霉素抗性基因的重组质粒pBZ 692,通过基因载体pGV 3850,我们将基因1和基因2引入了高等植物。结果证明基因1和基因2能促使烟草、向日葵、土豆等转化组织分化长根,转化的根在MS_0培养基上能脱分化形成愈伤组织并自主生长,在转化的组织中有转化标记胭脂碱的存在。     

A simple purification of calmodulin by reversed phase chromatography with hydrophobic polymer 3520

A new adsorption chromatography procedure for the purification of calmodulin from bovine brain was developed using polymeric adsorbent 3520. Calmodulin was first isolated by DEAE-Cellulose column chromatography and further purified to apparent homogeneity following elution with 50% ethanol from the adsorbent column. Polyacrylamide gel electrophoresis showed one band either in the presence of Ca2+ or EGTA. The polymeric adsorbent 3520 is a non-polar polymer lacking exchangeable groups. The selective adsorption of calmodulin is based on hydrophobic interaction within the matrix, and is Ca2+ independent. Neither high salt (0.5 M NaC1) nor EGTA (5 mM) was able to elute the CaM from the adsorption column whereas ethanol (50%) eluted it completely. This method is simple to use and it provides highly purified calmodulin with high yield.     

Primary structure of PDC-109, a major protein constituent of bovine seminal plasma

The two major protein components of bovine seminal plasma, PDC-109 and BSP I, have been purified by gel filtration, partition chromatography and reverse-phase high performance liquid chromatography from an 86% ethanol precipitate of bovine seminal plasma ejaculate. The complete 109-residue amino acid sequence of PDC-109 has been established by automated Edman degradation of the intact peptide as well as its proteolytic digestion and cyanogen bromide cleavage fragments. The 12,774 dalton structure has two structurally similar domains of 38 and 41 amino acids, each containing two disulfide bonds.     

Characterization and Location of Met5-Enkephalin-Arg6-Phe7 Stored in Various Rat Brain Regions

A specific antiserum against met⁵-enkepha-lin-arg⁶-phe⁷ was raised and used to study the distribution and characterization of met⁵-enkephalin-arg⁶-phe⁷-like immunoreactive material in rat brains by radioimmunoassay and immunohistochemical procedures. The antiserum appears to be directed to the COOH-terminus of the peptide, as it fails to cross-react with met⁵-enkeph-alin, met³-enkephalin-arg⁶, met⁵-enkephalin-arg⁶-arg⁷, met⁶-enkephalin-lys⁶, and leu-enkephalin. However, it cross-reacts with phe-met-arg-phe by about 10% and with phe-met-arg-phe-NH₂ to an insignificant degree. The highest content of met⁵-enkephalin-arg⁶-phe⁷ was found in the striatum, which contains a dense network of immunoreactive varicose fibers and terminals, as well as immunoreac tive cell bodies. The met⁵-enkephalin-arg⁶-phe⁷ in striatum can be released in a Ca²⁺-dependent manner by a depolarizing concentration of KC1, raising the possibility of a neu-roregulatory role for met⁵-enkephalin-arg⁶-phe⁷. Characterization of the immunoreactive material by gel filtration and high pressure liquid chromatography revealed the presence of multiple forms of immunoreactive material in some brain regions.     

Activity of central neurons of the electroreceptor system ofGlyptosternum reticulatum

Two types of electroreceptive neurons — tonic and phasic — were found in acute experiments with extracellular recording of unit activity from the lateral lobes of the medulla in the Turkestan catfishGlyptosternum reticulatum. Tonic neurons were more sensitive to the potential gradient in water (the threshold for most neurons was 1–6 µV/cm) than phasic neurons, they possessed spontaneous activity (mean frequency 4–10 spikes/sec), and their response characteristics depended significantly on the intensity and duration of stimulation. Phasic neurons had no spontaneous activity; their sensitivity was about one order of magnitude lower than that of the tonic neurons, and the response was independent of the parameters of the stimuli. The probable mechanisms of differentiation of neurons into two types, with a possible link with the characteristics of the receptor formations or with the functional organization of the corresponding brain centers, are discussed.     

Kinetics of the sodium current through EDTA-modified calcium channels of the molluscan neuron somatic membrane

The kinetics of the slow current carried by sodium ions through potential-dependent calcium channels after addition of EDTA to calcium-free external solution was investigated in experiments by the intracellular dialysis method on isolatedHelix pomatia neurons. The activation kinetics of this current was similar to that of the calcium current and could be described by the use of the square of the activation variable m in Hodgkin-Huxley equations. The decay (inactivation) kinetics of the induced sodium current during prolonged depolarization is biexponential in character. It is suggested that decay of the sodium currents takes place as a result of two independent processes: potential-dependent inactivation with a time constant τ_h～1 sec, taking place as far as a certain steady-state level h_∞, and a decrease in current connected with Na⁺ accumulation inside the cell during passage of the current and a consequent change in the sodium electrochemical potential (τ_c～10 sec). It is concluded that modification of the calcium channels, so that they acquire the ability to conduct sodium, has no significant effect on the gating mechanisms responsible for opening and closing of the channels.     

Serum Albumin Washing Specifically Enhances Arachidonate Incorporation into Synaptosomal Phosphatidylinositols

Arachidonate incorporation into synaptosomal phospholipids was shown to be affected by factors including the procedure for preparation of the membrane fractions and preincubation of synaptosomes prior to assay of incorporation of arachidonate into both phosphatidylcholine (PC) and phosphatidylinositol (PI). However, the inhibition toward incorporation into PIs, but not PCs, was fully reversed when the membranes were washed with bovine serum albumin. A twofold increase in arachidonate incorporation into PIs was also observed when freshly prepared synaptosomes were washed with serum albumin immediately before assay of incorporation activity. The inhibitory action is thought to be due to an increase in polyunsaturated fatty acids and/or their oxidation products which may then elicit a special effect on the acyltransferase responsible for transferring arachidonate into phosphatidylinositols. The differences in fatty acid uptake and response to serum albumin also suggest the presence of different acyltransferase for acyl transfer to PIs and PCs.     

Highly purified particulate guanylate cyclase from rat lung: characterization and comparison with soluble guanylate cyclase

Guanylate cyclase was purified 1000-fold from washed rat lung particulate fractions to a final specific activity of 500 nmoles cyclic GMP produced/min/mg protein by a combination of detergent extraction and chromatography on concanavalin A-Sepharose, GTP-agarose, and blue agarose. Particulate guanylate cyclase has a molecular weight of 200 000 daltons, a Stokes radius of 48 A and a sedimentation coefficient of 9.4 while the soluble form has a molecular weight of 150 000 daltons, a Stokes radius of 44 A, and a sedimentation coefficient of 7.0. Whereas the particulate enzyme is a glycoprotein with a specific affinity for concanavalin A and wheat germ agglutinin, the soluble form of guanylate cyclase did not bind to these lectins. Purified particulate guanylate cyclase did not cross-react with a number of monoclonal antibodies generated to the soluble enzyme. While both forms of the enzyme could be regulated by the formation of mixed disulfides, the particulate enzyme was relatively insensitive to inhibition by cystine. With GTP as substrate both forms of the enzyme demonstrated typical kinetics, and with GTP analogues negative cooperativity was observed with both enzyme forms. These data support the suggestion that the two forms of guanylate cyclase possess similar catalytic sites, although their remaining structure is divergent, resulting in differences in subcellular distribution, physical characteristics, and antigenicity.     

Adaptive Functions of Vertebrate Molting Cycles

SYNOPSIS Cyclic epidermal cellular prohfeiation,with or withoutkeratinization is a vertebrate characteristic Such activityprobably obeys an autonomous rhythm which is legulated throughneuro humoral S)stcms in response to envnonmental (piox imate)stimuliand related to adaptive (ultimate) factors In seeking causeand effect lelationships, however, it becomes apparent thatthe same environmental parameter may be both an ultimate anda pioximate factor, the latter also regulating the rate of lesponseWith regard to molting in homoio'heims, tempeiatuie acts insuch a capacity in many species Peiiodic shedding of the outer epidermis in fish amphibiansand reptiles does not appear to be coirelated with seasonalfactors to the extent that avian and mammalian molts are The evolution of vertebrate molting cycles has amounted to theentraining of inherent epidermal C)cles with seasonal demandsby the organism itself and the environment,these demands actas regulating mechanisms Pieadapted structures such as feathersand hairs function collectively as plumage and pelage in theirvarious roles but separately in their growth and leplacementcycles which, however, are coordinated for maximum functionalefficiency Molting is also synchionized with the seasonal cycleaccording to the availability of energy resources and time tocomplete the essential functions (in addition to molting) Theevolved molting systems as manifested in the gieat variety ofpatterns and types in the vertebrates, may thus be legardedas almost individual responses to selective piessures actingon a umveisil vertebrate chaiacter The basic regulatoiy system involves the neuro hvpophyseal complexwhich contiols target endocrines affecting various functionswhich themselves influence epidermal mitosis and, ultimately,molting 1 he mechanism in its simplest form controls the animalsmetabolism through the thyroid acting independently in a permissivecapacity or synergistically with the adrenal and gonadal hormoneswhich are regulated directly and/or indirectly through negativefeedback