Transient expression of a human ?-actin promoter/lacZ gene introduced into mouse embryos correlates with a low degree of methylation

1993 ◽  
Vol 34 (2) ◽  
pp. 149-157 ◽  
Author(s):  
Erik Nilsson ◽  
Urban Lendahl
Keyword(s):  
Transient Expression ◽  
Mouse Embryos ◽  
Lacz Gene ◽  
Low Degree ◽  
Actin Promoter

scholarly journals Agrobacterium-mediated gene delivery and transient expression in the red macroalga Chondrus crispus

2018 ◽  
Vol 61 (5) ◽  
pp. 499-510 ◽  
Author(s):  
Anusha Devi Ramessur ◽  
John H. Bothwell ◽  
Christine A. Maggs ◽  
Sook Yee Gan ◽  
Siew Moi Phang
Keyword(s):  
Transient Expression ◽  
Red Algae ◽  
Transformation Efficiency ◽  
Bacterial Cells ◽  
Chondrus Crispus ◽  
Cultivation Medium ◽  
Intimate Association ◽  
Actin Promoter ◽  
Transfer Method ◽  
Gene Transfer Method

Abstract Molecular resources and transgenic studies in red algae are lagging behind those for green algae. The Agrobacterium-mediated gene-transfer method routinely used in plant transformation has not been fully utilised in the red algae, which, as an important source of phycocolloids, warrant more studies. In this regard, a stepwise methodology was developed for Agrobacterium-mediated transformation of the carrageenophyte Chondrus crispus using pCAMBIA 1301 and a construct featuring a codon-optimized beta-glucuronidase (GUS) reporter gene driven by the endogenous Chondrus actin promoter. The effects of several factors on transformation efficiency were investigated. An intimate association of Chondrus and bacterial cells was observed using scanning electron microscopy. GUS transient expression within Chondrus cortical and medullary cells with both expression cassettes testified to the amenability of Chondrus to Agrobacterium-mediated transformation. Darker staining, indicative of higher GUS activity, was observed with the Chondrus-specific construct, suggesting its superiority over the pCAMBIA 1301. Presence of acetosyringone, the wounding method and the type of co-cultivation medium significantly affected the transformation outcome and efficiency. The Agrobacterium-mediated transient expression presented here constitutes a first step towards tailoring a transformation strategy for Chondrus, which can serve to facilitate further transgenic studies in this important red alga.

In vitro manipulation of early mouse embryos induces HIV1-LTRlacZ transgene expression

Development ◽  
1993 ◽  
Vol 119 (4) ◽  
pp. 1293-1300 ◽  
Author(s):  
M. Vernet ◽  
C. Cavard ◽  
A. Zider ◽  
P. Fergelot ◽  
G. Grimber ◽  
...  
Keyword(s):  
Transgene Expression ◽  
Molecular Mechanisms ◽  
Constitutive Expression ◽  
Mouse Embryos ◽  
Lacz Gene ◽  
Cell Stage ◽  
Early Mouse ◽  
Transgenic Embryos

We report here that the transcriptional activity of early mouse embryos is affected by their manipulation and culture in vitro, using transgenic embryos that express the reporter gene lacZ. We examined the pattern of expression of the lacZ gene fused to the human immunodeficiency virus type 1 long terminal repeat during the preimplantation stages. Transgene expression is induced as early as the two-cell stage in embryos developed in vitro, while there is no constitutive expression at the same stage in embryos developed in vivo. We have established a relation between this inducible expression occurring in vitro and an oxidative stress phenomenon. Indeed, when the culture medium is supplemented with antioxidants such N-acetyl-cysteine or CuZn-superoxide dismutase the transgene expression is markedly reduced. We also present evidence that the transgene expression in vitro coincides with the onset of the embryonic genome activation as attested by the synthesis of the 70 × 10(3) M(r) protein complex. Therefore, this transgene expression could prove to be a useful tool in our understanding of the molecular mechanisms involved in this crucial developmental event.

Human Adenovirus Uptake by Preirnplantation Mouse Embryos

1972 ◽  
Vol 30 ◽  
pp. 268-269
Author(s):  
D. G. Chase ◽  
W. Winters ◽  
L. Piko
Keyword(s):  
Hela Cells ◽  
Human Adenovirus ◽  
Mouse Embryos ◽  
Equilibrium Density ◽  
Cell Stage ◽  
Virus Attachment ◽  
Preimplantation Mouse Embryos ◽  
Embryo Culture Medium ◽  
Preimplantation Mouse ◽  
Mouse Cells

Although the outlines of human adenovirus entry and uncoating in HeLa cells has been clarified in recent electron microscope studies, several details remain unclear or controversial. Furthermore, morphological features of early interactions of human adenovirus with non-permissive mouse cells have not been extensively documented. In the course of studies on the effects of human adenoviruses type 5 (AD-5) and type 12 on cultured preimplantation mouse embryos we have examined virus attachment, entry and uncoating. Here we present the ultrastructural findings for AD-5.AD-5 was grown in HeLa cells and purified by successive velocity gradient and equilibrium density gradient centrifugations in CsCl. After dialysis against PBS, virus was sedimented and resuspended in embryo culture medium. Embryos were placed in culture at the 2-cell stage in Brinster's medium.

Preplate neurons in embryonic mouse cortex: Ultrastructural observations using photoconversion of the fluorescent lipophilic dye dil

1992 ◽  
Vol 50 (1) ◽  
pp. 906-907
Author(s):  
Linda C. Hassinger ◽  
James E. Crandall
Keyword(s):  
Mouse Embryos ◽  
Embryonic Mouse ◽  
Cortical Afferents ◽  
Mouse Cortex ◽  
Carbocyanine Dye ◽  
Increased Sensitivity

We have begun to look directly at small numbers of afferent axons to early generated neurons that form the preplate in the developing mouse cortex. The carbocyanine dye Dil (1’1, dioctadecyl-3,3,3’3’-tetramethyl-indocarbocyanine) has proved especially useful for this goal. DiI labels axons and their terminals with greater sensitivity and without some of the disadvantages of axon filling with HRP. The increased sensitivity provided by labeling embryonic axons with DiI has given us new insights into the development of cortical afferents. For instance, we reported originally that afferents from the thalamus were present below the cortex as early as embryonic day 15 (E15) based on HRP injections into mouse embryos. By using DiI placements into the thalamus in aldehyde-fixed brains, we now know that thalamic fibers reach the cortex 24 hrs earlier.

Localization of Intracisternal a Particle Antigens in Neoplastic Cells and Preimplantation Mouse Embryos

1980 ◽  
Vol 38 ◽  
pp. 696-697
Author(s):  
Thomas T.F. Huang ◽  
Patricia G. Calarco
Keyword(s):  
Endoplasmic Reticulum ◽  
Normal Development ◽  
Mouse Embryos ◽  
Neoplastic Cells ◽  
Large Numbers ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse ◽  
Intracisternal A Particle ◽  
Normal Feature

The stage specific appearance of a retravirus, termed the Intracisternal A particle (IAP) is a normal feature of early preimplantation development. To date, all feral and laboratory strains of Mus musculus and even Asian species such as Mus cervicolor and Mus pahari express the particles during the 2-8 cell stages. IAP form by budding into the endoplasmic reticulum and appear singly or as groups of donut-shaped particles within the cisternae (fig. 1). IAP are also produced in large numbers in several neoplastic cells such as certain plasmacytomas and rhabdomyosarcomas. The role of IAP, either in normal development or in neoplastic behavior, is unknown.

Assessment of Damage to Human Platelets after Aggregation and Other Injuries by Microscopic Observation and Estimation of Serotonin Uptake

1970 ◽  
Vol 23 (02) ◽  
pp. 261-275 ◽  
Author(s):  
G Zbinden ◽  
J. N Mehrishi ◽  
S Tomlin
Keyword(s):  
Platelet Aggregation ◽  
Electrophoretic Mobility ◽  
Human Platelets ◽  
Freezing And Thawing ◽  
Low Degree ◽  
Microscopic Evaluation ◽  
Negative Surface ◽  
Negative Surface Charge ◽  
Charged Macromolecules ◽  
5 Ht Uptake

SummaryThe severity of platelet damage induced by hyper- and hypotonic NaCl solutions and freezing and thawing was assessed by microscopic evaluation and measuring inhibition of 5-HT uptake. The same techniques were used to quantitate the effects of aggregating agents. The positively charged macromolecules PS, Poly-L und Poly-O reduced the net negative surface charge as determined by microelectrophoresis, caused platelet aggregation and inhibited 5-HT uptake. The damaging effects of Poly-L and Poly-O were more severe and more closely related to concentration than that of PS. The negatively charged macromolecules Poly-IC and NaPS increased the anodic electrophoretic mobility. Poly-IC and heparin caused a low degree of platelet clumping and no inhibition of 5-HT uptake. NaPS produced severe platelet damage with extensive clumping and complete inhibition of 5-HT uptake. Na laurate had the same effect, but did not alter electrophoretic mobility. ADP caused concentration-dependent platelet aggregation and inhibition of 5-HT uptake. The effects of ADP and NaPS were compared in agitated and non-agitated platelet samples containing identical concentrations of the 2 compounds. Agitation was found to increase the degree of platelet clumping and to reduce 5-HT uptake.

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