We have constructed a pTZ19R-derived vector which allows efficient positive selection of recombinant plasmids. The system uses the EcoK restriction activity of Escherichia coli K-12 to select against non-recombinant plasmids. The vector contains an EcoK site which, if deleted or disrupted by ligating a DNA fragment, yields recombinant plasmids that are no longer susceptible to EcoK restriction when transformed into a restriction-proficient E. coli host.
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http://dx.doi.org/10.1016/0378-1119(94)90885-0 | DOI Listing |
Research (Wash D C)
January 2025
Cancer Institute, Xuzhou Medical University, Xuzhou, Jiangsu 221004, China.
Chimeric antigen receptor (CAR)-based immune cell therapies attack neighboring cancer cells after receptor recognition but are unable to directly affect distant tumor cells. This limitation may contribute to their inefficiency in treating solid tumors, given the restricted intratumoral infiltration and immunosuppressive tumor microenvironment. Therefore, cell-cell fusion as a cell-killing mechanism might develop a novel cytotherapy aimed at improving the efficacy against solid tumors.
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January 2025
Department of Biology, School of Sciences and Humanities, Nazarbayev University, Astana, Kazakhstan.
Background: Chitosan nanoparticles (CsNPs) are an effective and inexpensive approach for DNA delivery into live cells. However, most CsNP synthesis protocols are not optimized to allow long-term storage of CsNPs without loss of function. Here, we describe a protocol for CsNP synthesis, lyophilization, and sonication, to store CsNPs and maintain transfection efficiency.
View Article and Find Full Text PDFACS Synth Biol
January 2025
Department of Life Sciences, Imperial College London, London SW7 2AZ, U.K.
Naturally occurring DNA inversion systems play an important role in the generation of genetic variation and adaptation in prokaryotes. Shufflon invertase (SI) from plasmid R64, recognizing asymmetric sites, has been adopted as a tool for synthetic biology. However, the availability of a single enzyme with moderate rates of recombination has hampered the more widespread use of SIs.
View Article and Find Full Text PDFZhonghua Yi Xue Za Zhi
February 2025
Department of Orthopedics, the First Hospital of Huaian City, Nanjing Medical University, Huaian 223300, China.
To investigate the effects of long non-coding RNA KLHL7-AS1 (LncRNA KLHL7-AS1) on the proliferation and apoptosis of nucleus pulposus cells under oxidative stress and its mechanisms. Human nucleus pulposus cells (HUM-iCell-s012) were divided into 4 groups, and unoxidized nucleus pulposus cells were transfected with an empty pcDNA vector (pcDNA-control) to serve as the blank control group. Based on previous studies on oxidative stress-induced nucleus pulposus cell senescence and preliminary experiments, oxidative stress was induced by treating nucleus pulposus cells with 400 μmol/L HO.
View Article and Find Full Text PDFFish Shellfish Immunol
January 2025
Tianjin Key Lab of Aqua-Ecology and Aquaculture, College of Fisheries, Tianjin Agricultural University, Tianjin, 300392, China. Electronic address:
Rahnella aquatilis is an emerging opportunistic pathogen that usually causes septicaemia in fish and poses a potential threat to human health. VgrG gene is an important virulence factor of type VI secretion system in R. aquatilis, but its regulatory mechanism underlying PANoptosis is still unknown.
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