scholarly journals A new kinesin-like protein (Klp1) localized to a single microtubule of the Chlamydomonas flagellum.

1994 ◽  
Vol 125 (6) ◽  
pp. 1313-1326 ◽  
Author(s):  
M Bernstein ◽  
P L Beech ◽  
S G Katz ◽  
J L Rosenbaum

The kinesin superfamily of mechanochemical proteins has been implicated in a wide variety of cellular processes. We have begun studies of kinesins in the unicellular biflagellate alga, Chlamydomonas reinhardtii. A full-length cDNA, KLP1, has been cloned and sequenced, and found to encode a new member of the kinesin superfamily. An antibody was raised against the nonconserved tail region of the Klp1 protein, and it was used to probe for Klp1 in extracts of isolated flagella and in situ. Immunofluorescence of whole cells indicated that Klp1 was present in both the flagella and cell bodies. In wild-type flagella, Klp1 was found tightly to the axoneme; immunogold labeling of wild-type axonemal whole mounts showed that Klp1 was restricted to one of the two central pair microtubules at the core of the axoneme. Klp1 was absent from the flagella of mutants lacking the central pair microtubules, but was present in mutant flagella from pf16 cells, which contain an unstable C1 microtubule, indicating that Klp1 was bound to the C2 central pair microtubule. Localization of Klp1 to the C2 microtubule was confirmed by immunogold labeling of negatively stained and thin-sectioned axonemes. These findings suggest that Klp1 may play a role in rotation or twisting of the central pair microtubules.

1985 ◽  
Vol 5 (3) ◽  
pp. 569-575
Author(s):  
R Sperling ◽  
J Sperling ◽  
A D Levine ◽  
P Spann ◽  
G R Stark ◽  
...  

Transcripts of the CAD gene in Syrian hamster cells are as abundant in the nucleus as in the cytoplasm. This was shown by in situ hybridization of whole cells and by solution and blot hybridization of subcellular fractions. Similar results were obtained both for wild-type cells and for a mutant containing amplified CAD genes in which the level of CAD RNA is 150-fold greater. CAD nuclear RNA is indistinguishable from mature mRNA by gel electrophoresis and blot hybridization. Discrete higher-molecular-weight precursors are undetectable, although the persistence of a short length of intervening sequence in the otherwise fully processed RNA is not excluded. CAD RNA is released from nuclei by sonication in physiological conditions in a ribonucleoprotein form that sediments as a broad peak at about 200S in a sucrose gradient. CAD sequences extracted from nuclei by treatment with EDTA and RNase are found in the 30S particles previously described.


2009 ◽  
Vol 30 (2) ◽  
pp. 382-398 ◽  
Author(s):  
Mary Ann Checkley ◽  
Kunio Nagashima ◽  
Stephen J. Lockett ◽  
Katherine M. Nyswaner ◽  
David J. Garfinkel

ABSTRACT Ty1 is a retrovirus-like retrotransposon whose replication is influenced by diverse cellular processes in Saccharomyces cerevisiae. We have identified cytoplasmic P-body components encoded by DHH1, KEM1, LSM1, and PAT1 as cofactors that posttranscriptionally enhance Ty1 retrotransposition. Using fluorescent in situ hybridization and immunofluorescence microscopy, we found that Ty1 mRNA and Gag colocalize to discrete cytoplasmic foci in wild-type cells. These foci, which are distinct from P-bodies, do not form in P-body component mutants or under conditions suboptimal for retrotransposition. Our immunoelectron microscopy (IEM) data suggest that mRNA/Gag foci are sites where virus-like particles (VLPs) cluster. Overexpression of Ty1 leads to a large increase in retrotransposition in wild-type cells, which allows VLPs to be detected by IEM. However, retrotransposition is still reduced in P-body component mutants under these conditions. Moreover, the percentage of Ty1 mRNA/Gag foci and VLP clusters and levels of integrase and reverse transcriptase are reduced in these mutants. Ty1 antisense RNAs, which have been reported to inhibit Ty1 transposition, are more abundant in the kem1Δ mutant and colocalize with Ty1 mRNA in the cytoplasm. Therefore, Kem1p may prevent the aggregation of Ty1 antisense and mRNAs. Overall, our results suggest that P-body components enhance the formation of retrotransposition-competent Ty1 VLPs.


1985 ◽  
Vol 5 (3) ◽  
pp. 569-575 ◽  
Author(s):  
R Sperling ◽  
J Sperling ◽  
A D Levine ◽  
P Spann ◽  
G R Stark ◽  
...  

Transcripts of the CAD gene in Syrian hamster cells are as abundant in the nucleus as in the cytoplasm. This was shown by in situ hybridization of whole cells and by solution and blot hybridization of subcellular fractions. Similar results were obtained both for wild-type cells and for a mutant containing amplified CAD genes in which the level of CAD RNA is 150-fold greater. CAD nuclear RNA is indistinguishable from mature mRNA by gel electrophoresis and blot hybridization. Discrete higher-molecular-weight precursors are undetectable, although the persistence of a short length of intervening sequence in the otherwise fully processed RNA is not excluded. CAD RNA is released from nuclei by sonication in physiological conditions in a ribonucleoprotein form that sediments as a broad peak at about 200S in a sucrose gradient. CAD sequences extracted from nuclei by treatment with EDTA and RNase are found in the 30S particles previously described.


Author(s):  
B.A. Hamkalo ◽  
S. Narayanswami ◽  
A.P. Kausch

The availability of nonradioactive methods to label nucleic acids an the resultant rapid and greater sensitivity of detection has catapulted the technique of in situ hybridization to become the method of choice to locate of specific DNA and RNA sequences on chromosomes and in whole cells in cytological preparations in many areas of biology. It is being applied to problems of fundamental interest to basic cell and molecular biologists such as the organization of the interphase nucleus in the context of putative functional domains; it is making major contributions to genome mapping efforts; and it is being applied to the analysis of clinical specimens. Although fluorescence detection of nucleic acid hybrids is routinely used, certain questions require greater resolution. For example, very closely linked sequences may not be separable using fluorescence; the precise location of sequences with respect to chromosome structures may be below the resolution of light microscopy(LM); and the relative positions of sequences on very small chromosomes may not be feasible.


Author(s):  
Juan Alfredo Lino-Gamiño ◽  
Carlos Méndez-González ◽  
Eduardo José Salazar-Araujo ◽  
Pablo Adrián Magaña-Sánchez

In the value chain it is important to keep in mind the core business of the company, since it depends largely on the competitiveness of the company and its overall performance, bearing in mind that all business indicators depend on it. In this work we will study the washing process within the company WASH CONTAINERS SA DE CV, to improve the washing processes and in this way reduce times and movements in the process leading the company to reduce costs considerably within the operations company daily, having a more competitive operation and with greater profit margin in its business process. Goals: It Improve the logistics of the movement of containers for washing and with it the core business of the company. Methodology: The action research will be applied applying Business Process Management for the improvement of processes in situ, it will be developed in a certain period of time and with that it will establish an improvement projection. Contribution: The improvement of the times for the disposal of the containers and their subsequent use, allows a better competitiveness and with it the income of the company, on the other hand, the transport companies improve in performance in quantity, quality of disposition and with it their income.


Pathogens ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 54
Author(s):  
Christine Landlinger ◽  
Lenka Tisakova ◽  
Vera Oberbauer ◽  
Timo Schwebs ◽  
Abbas Muhammad ◽  
...  

Bacterial vaginosis is characterized by an imbalance of the vaginal microbiome and a characteristic biofilm formed on the vaginal epithelium, which is initiated and dominated by Gardnerella bacteria, and is frequently refractory to antibiotic treatment. We investigated endolysins of the type 1,4-beta-N-acetylmuramidase encoded on Gardnerella prophages as an alternative treatment. When recombinantly expressed, these proteins demonstrated strong bactericidal activity against four different Gardnerella species. By domain shuffling, we generated several engineered endolysins with 10-fold higher bactericidal activity than any wild-type enzyme. When tested against a panel of 20 Gardnerella strains, the most active endolysin, called PM-477, showed minimum inhibitory concentrations of 0.13–8 µg/mL. PM-477 had no effect on beneficial lactobacilli or other species of vaginal bacteria. Furthermore, the efficacy of PM-477 was tested by fluorescence in situ hybridization on vaginal samples of fifteen patients with either first time or recurring bacterial vaginosis. In thirteen cases, PM-477 killed the Gardnerella bacteria and physically dissolved the biofilms without affecting the remaining vaginal microbiome. The high selectivity and effectiveness in eliminating Gardnerella, both in cultures of isolated strains as well as in clinically derived samples of natural polymicrobial biofilms, makes PM-477 a promising alternative to antibiotics for the treatment of bacterial vaginosis, especially in patients with frequent recurrence.


2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Martin Edwards ◽  
Pierre Hélaouët ◽  
Eric Goberville ◽  
Alistair Lindley ◽  
Geraint A. Tarling ◽  
...  

AbstractIn the North Atlantic, euphausiids (krill) form a major link between primary production and predators including commercially exploited fish. This basin is warming very rapidly, with species expected to shift northwards following their thermal tolerances. Here we show, however, that there has been a 50% decline in surface krill abundance over the last 60 years that occurred in situ, with no associated range shift. While we relate these changes to the warming climate, our study is the first to document an in situ squeeze on living space within this system. The warmer isotherms are shifting measurably northwards but cooler isotherms have remained relatively static, stalled by the subpolar fronts in the NW Atlantic. Consequently the two temperatures defining the core of krill distribution (7–13 °C) were 8° of latitude apart 60 years ago but are presently only 4° apart. Over the 60 year period the core latitudinal distribution of euphausiids has remained relatively stable so a ‘habitat squeeze’, with loss of 4° of latitude in living space, could explain the decline in krill. This highlights that, as the temperature warms, not all species can track isotherms and shift northward at the same rate with both losers and winners emerging under the ‘Atlantification’ of the sub-Arctic.


Catalysts ◽  
2019 ◽  
Vol 9 (10) ◽  
pp. 792 ◽  
Author(s):  
Andrés R. Alcántara

Biocatalysis is the term used to describe the application of any type of biocatalyst (enzymes, as isolated preparations of wild-type or genetically modified variants, or whole cells, either as native cells or as recombinant expressed proteins inside host cells) in a given synthetic schedule [...]


1998 ◽  
Vol 519 ◽  
Author(s):  
L. Bergogne ◽  
S. Fennouh ◽  
J. Livage ◽  
C. Roux

AbstractBioencapsulation in sol-gel materials has been widely studied during the past decade. Trapped species appear to retain their bioactivity in the porous silica matrix. Small analytes can diffuse through the pores allowing bioreactions to be performed in-situ, inside the sol-gel glass. A wide range of biomolecules and micro-organisms have been encapsulated. The catalytic activity of enzymes is used for the realization of biosensors or bioreactors. Antibody-antigen recognition has been shown to be feasible within sol-gel matrices. Trapped antibodies bind specifically the corresponding haptens and can be used for the detection of traces of chemicals. Even whole cells are now encapsulated without any alteration of their cellular organization. They can be used for the production of chemicals or as antigens for immunoassays.


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