3 research outputs found
A method for improving development potential of an embryo and embryos developed therefrom
The present invention relates to a method for improving development potential of an embryo, embryos developed therefrom and organisms resulting from embryos developed from the method
Silencing Transcription from an Influenza Reverse Genetics Plasmid in E. coli Enhances Gene Stability
Reverse genetics
(RG) systems have been instrumental for determining
the molecular aspects of viral replication, pathogenesis, and for
the development of therapeutics. Here, we demonstrate that genes encoding
the influenza surface antigens hemagglutinin and neuraminidase have
varying stability when cloned into a common RG plasmid and transformed
into Escherichia coli. Using GFP as
a reporter, we demonstrate that E. coli expresses the target genes in the RG plasmid at low levels. Incorporating lac operators or a transcriptional terminator into the plasmid
reduced expression and stabilized the viral genes to varying degrees.
Sandwiching the viral gene between two lac operators
provided the largest contribution to stability and we confirmed the
stabilization is Lac repressor-dependent and crucial for subsequent
plasmid propagations in E. coli. Viruses
rescued from the lac operator-stabilized plasmid
displayed similar kinetics and titers to the original plasmid in two
different viral backbones. Together, these results indicate that silencing
transcription from the plasmid in E. coli helps to maintain the correct influenza gene sequence and that the lac operator addition does not impair virus production.
It is envisaged that sandwiching DNA segments between lac operators can be used for reducing DNA segment instability in any
plasmid that is propagated in E. coli which express the Lac repressor
Poly(cannabinoid)s: Hemp-Derived Biocompatible Thermoplastic Polyesters with Inherent Antioxidant Properties
The
legalization of hemp cultivation in the United States has caused
the price of hemp-derived cannabinoids to decrease 10-fold within
2 years. Cannabidiol (CBD), one of many naturally occurring diols
found in hemp, can be purified in high yield for low cost, making
it an interesting candidate for polymer feedstock. In this study,
two polyesters were synthesized from the condensation of either CBD
or cannabigerol (CBG) with adipoyl chloride. Poly(CBD-Adipate) was
cast into free-standing films and subjected to thermal, mechanical,
and biological characterization. Poly(CBD-Adipate) films exhibited
a lack of cytotoxicity toward adipose-derived stem cells while displaying
an inherent antioxidant activity compared to poly(lactide) films.
Additionally, this material was found to be semi-crystalline and able
to be melt-processed into a plastic hemp leaf using a silicone baking
mold