Restriction map of Tn7

Tn7, a transposon of 14 kb, encodes resistance to trimethoprim (Tp) and streptomycin (Sm). A cleavage site map of this transposon for twenty-two different restriction enzymes as determined by comparison of restriction enzyme cleavage patterns of the plasmids ColE1 and ColE1::Tn7 is presented. The precise localization of these sites was facilitated by the use of two deletion derivatives of ColE1::Tn7: pGB2 and ColE1::Tn7Δ6, and by the use of pOB14 and pOB15 which contain a part of Tn7 cloned into the plasmid pBR322. This map should aid in the study of the structural and genetic organization of this transposon.     

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     

Quantum theory of nerve excitation

Based on quantum transitions of membrane dipoles, the four fundamental properties of nerve impulse are derived in this paper: the all-or-none response, the strength-duration relation, refractoriness and refractory period and frequency modulation. Furthermore, the theory offers a physical mechanism for nerve excitation similar to a two-level ammonia maser. It also implies non-threshold excitation at elevated temperatures. The role of trimethylamine ions near the surface of a phospholipid membrane is briefly discussed to indicate a possible connection between theory and reality.     

Recombinants between Clock Mutants of CHLAMYDOMONAS REINHARDI

Mutants affecting the period length of the biological clock in Chlamydomonas reinhardi have been isolated and a start has been made on analyzing the genetics of this system. In four mutants, the long period characteristic seems to be controlled by single genes at separate loci. Crosses between single mutants, as well as crosses involving three or four mutant genes, yielded progeny with periods characteristic of the parents as well as recombinant types, including normal period (wild type) and extra-long periods (double, triple and quadruple mutants). It was found that the period lengthening effect is additive; that is, the period of double mutants is lengthened by the sum of the period lengthening of the single mutants.     

In Vitro Effect of Rifampin on Mycobacteria

Rifampin inhibited 20 strains of Mycobacterium tuberculosis in concentrations of 0.005 to 0.02 mug/ml in 7H-9 broth with Tween 80 and killed all or nearly all of the inoculum in four to eight times greater concentrations. In the same medium without Tween 80, as well as on 7H-10 agar, about 16 to 64 times these amounts were required to produce the same effect. Rifampin was also active against M. kansasii and some of the nonchromogenic mycobacteria. The incidence of mycobacterial cells resistant to rifampin within the cultures studied was in the range of one to four per 10(8) to 10(9) colony-forming units with concentrations of 4 to 125 mug of rifampin per ml. Only one of the Battey cultures and that of M. fortuitum yielded cells resistant to rifampin at 125 mug/ml but not at 500 mug/ml. The same strains yielded more than double that number of organisms resistant to streptomycin and up to 100 times more organisms resistant to isoniazid. All three drugs stopped the growth or reduced the mycobacterial population in growing cultures after contact for 24 to 48 hr. Complete inhibition of growth was produced by rifampin at 1.0 mug/ml in an average of 6 days and by streptomycin at 5.0 mug/ml in 3 days. After an average contact of 10.7 days with rifampin, five of seven strains resumed growth and all strains began regrowth after exposure to streptomycin for 9.4 days. The marked susceptibility of M. tuberculosis and of atypical mycobacteria to rifampin in vitro and the relatively low incidence of resistant mutants suggests that this agent may have clinical usefulness in the treatment of tuberculosis and some other mycobacterioses.     

Bacterial Metabolism of Mevalonic Acid

Soluble cell-free extracts of actinomycete S4 grown on media containing mevalonate catalyze acetoacetate formation from mevalonate, mevaldate, and β-hydroxy-β-methylglutaryl-coenzyme A (CoA). Conversion of mevalonate to acetoacetate involves formation of free β-hydroxy-β-methylglutaryl-CoA, but not free mevaldate. The reaction favors mevalonate oxidation, and nicotinamide adenine dinucleotide, rather than nicotinamide adenine dinucleotide phosphate, acts as oxidant.