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1.
Several unit-length minicircles from the kinetoplast DNA of Leishmania tarentolae were cloned into pBR322 and into M13 phage vectors. The complete nucleotide sequences of three different partially homologous minicircles were obtained. The molecules contained a region of approx. 80% sequence homology extending for 160–270 bp and a region unique to each minicircle. A 14-mer was found to be conserved in all kinetoplast minicircle sequences reported to date. The frequency distributions of various minicircle sequence classes in L. tarentolae were obtained by quantitative gel electrophoresis and by examination of the “T ladder” patterns of minicircles randomly cloned into M13 at several sites. By these methods we could assign approx. 50% of the total minicircle DNA into a minimum of five sequence classes. A sequence-dependent polyacrylamide gel migration abnormality was observed with several minicircle fragments both cloned and uncloned. The abnormality was dependent on the presence of a portion of the conserved region of the minicircle. 相似文献
2.
In this paper, we describe an efficient procedure for the purification of yeast phosphofructokinase. This procedure eliminates any time delay and enables to obtain an enzyme with minimum proteolytic alterations. The molecular weights of the oligomeric enzyme and of its constitutive subunits were both evaluated by means of several independent methods. However, the accuracy of each measurement was not sufficient to discriminate between an hexameric and an octameric structure of the enzyme oligomer. On the other hand, crosslinking experiments demonstrated the octameric structure of yeast phosphofructokinase. Obviously, some methods of molecular weight determination have led to erroneous results. In particular, our experiments show that the reliability of molecular weight determinations performed by gel filtration of native proteins must be considered with caution. 相似文献
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4.
Summary Individuals of the two-spotted spider mite, Tetranychus urticae Koch (Acarina: Tetranychidae), with and without a capacity for diapause, coexist in central Japan. Diapause appears to be adaptive because females in diapause suffered less mortality than non-diapausing individuals when frozen at –24°C for more than 4 h. However non-diapausing females showed good survival up to 4 h of freezing. Active non-diapausing mites survived on rose leaves in Kyoto (35°N) throughout the winters of 1988–1989 and 1989–1990. Cultures of mites with low (LD) and high (HD) diapause capacities at 18°C and 9L-15D photoperiod were successfully selected from the rose population and from a population on chrysanthemum in Nara (34.4°N). Their life-history traits at 15, 20, 25 and 30°C were characterized. HD and LD mites from both populations were of similar ages at first reproduction at 15–30° C. However, at temperatures 20° C, HD individuals produced more eggs than LD individuals, resulting in higher fecundity and intrinsic rate of natural increase. These traits allow HD individuals, which stop breeding between October and April, to increase faster in summer than LD individuals. This provides a mechanism, together with climatic fluctuations, in maintaining the coexistence of diapausing and non-diapausing T. urticae in Kyoto where winter conditions are rarely lethal to the non-diapausing individuals. 相似文献
5.
Claudio Nicolini Andrew S. Belmont Antonietta Martelli 《Cell biochemistry and biophysics》1986,8(2):103-117
Using HeLa S-3 cells synchronized by selective detachment, in this paper we report a parallel study of nuclear morphology
and autoradiography grain patterns between middle G1 and middle S phases: Our results show two distinct [3H]-thymidine labeling patterns. The first “peripheral” labeling pattern has a characteristic nuclear size distribution, in
contrast to the heterogeneous and varying size distributions of Feulgen-stained nuclei, and apparently is characteristic of
very early S phase. The sizes of the second labeling pattern—homogeneous or inhomogeneous grain distribution throughout the
nucleus—are equal or larger than the first and vary with S phase progression. Together, the corresponding nuclear sizes of
the labeled nuclei represent the larger extreme of nuclear areas, and the labeling index closely parallels the fraction of
nuclei with areas larger than the minimum size of the labeled nuclei. These results suggest a characteristic nuclear size
(reflecting unique intranuclear DNA distribution) as a necessary, if not sufficient, requirement for S phase initiation. Parallel
experimentation with rat liver cells—synchronized in vivo by partial hepatectomy and analyzed by thin section autoradiography—confirms
the existence of a peripheral labeling pattern in both the very early part and the very late part of S phase, which reconciles
our data with previous results and points to the fact that both initiation and termination sites for DNA replication are near
the nuclear periphery. 相似文献
6.
Francis Biville 《FEMS microbiology letters》1985,28(1):73-76
Abstract Pyrophosphate (PPi ), a noncompetitive inhibitor of Rho poly(C)-dependent ATPase activity in vitro has been shown to relieve polarity in the lac operon. This indicates that PPi could inhibit Rho activity in vivo too. An additional effect of PPi on adenosine 3',5'-cyclic monophosphate (cAMP) synthesis during stationary phase of growth is also described. 相似文献
7.
Bruna Tadolini Diana Fiorentini Laura Landi Luciana Cabrini 《Free radical research》1989,5(4):245-252
To find experimental conditions to selectively study the propagation phase of lipoperoxidation we studied the lipoperoxidation, catalyzed by FeCl2, of liposomes in a buffering condition where Fe2+ autoxidation and oxygen active species generation does not occur. Liposomes from egg yolk phosphatidylcholine. prepared by vortex mixing, do not oxidize Fe2+: on the contrary they oxidize Fe2+ when prepared by ultrasonic irradiation. Dimyristoyl phosphatidylcholine liposomes prepared by ultrasonic irradiation do not oxidize Fe2+. During sonication polyunsaturated fatty acid residues autoxidize and lipid hydroperoxides (LOOH) are generated. Only when LOOH are present in the liposimes Fe2+ oxidizes and its rate of oxidation depends on the amount of LOOH in the assay. The reaction results in the generation of both LOOH and thiobarbituric acid reactive material (TBAR): it is inhibited by butylated hydroxytoluene and has a acidic pH optimum; it is not inhibited by catalase and OH' scavengers. The reaction studied. thus, appears to be the chain branching and propagation phase of lipoperoxidation. When we studied the dependence of Fe2+ oxidation, LOOH and TBAR generation on FeCl2 concentration, we observed that at high FeCl2 concentrations the termination phase of lipoperoxidation was prevalent. Thus. by selecting the appropriate FeCl2 concentration the proposed experimental system allows study of either the propagation or the termination phase of lipoperoxidation. 相似文献
8.
1. | Under laboratory conditions complete development of H. sparsutum is shown within a temperature range of-0.7°C to +8°C. Constant temperatures above 10°C are lethal to the population. Larval growth (L1 to L5) is strictly temperature-dependent between-0.7 and +3.5°C, but slightly temperature compensated between +3.5 and +8°C. |
2. | Rate of egg production is highest at an average daily temperature of-0.7°C. |
3. | The sixth larval stage (L6) can be subdivided into a wandering and feeding period of about 40 days (at 7.3°C) and a following prepupal resting stage (PPR) with a high variability in duration, even at one and the same temperature, which ends with pupation. |
4. | Entrance into this resting stage seems to be independent of environmental changes and can occur at every time of the year. Termination of the PPR depends upon light stimuli received during the entire larval period. |
5. | Readiness for pupation exists when the photoperiod in the PPR exceeds that during hatching of first larva. A decreasing photoperiod during PPR triggers pupation in prepupae of various age. |
6. | The diapause stage seems to be a primary factor synchronizing the life cycle with seasonal changes in the environment. |
9.
10.
E.coli RNA聚合酶ββ'亚单位编码的基因rpoBC与核糖核蛋白基因rplJL共同构成rpoBC操纵子。rpl-rpo间区转录终止信号~tL7的转录产物RNA有两组富含C-G的反向对称结构及一串寡聚U;反向对称区可形成1:2和3:4茎环或单一5:6茎环。 用M13mp11噬菌体插入~tL7的112bp片段重组M13mp11-490,在此基础上用定点突变技术建立~tL7的七核苷酸缺失突变体,从而破坏1:2茎环,建成M13mp11-85。 分别把原型~tL7及缺失型~tL7~v克隆到表达质粒pHR24中,构建成pHR37(P_ftsQ~tL7-galk)及pHR24-9(p_ftsQ-~tL7~v-galk)。测定galk基因产物半乳糖激酶活性,推算转录的终止效率。结果表明:~tL7终止效率为50%,~tL7~v为20%。说明仅有3:4茎环及寡聚U,即具终止作用; 1:2茎环通过某种方式加强3:4茎环从而提高终止效率。 相似文献