Quantitative assessment on the cloning efficiencies of lentiviral transfer vectors with a unique clone site.
Gang Zhang,Anurag Tandon +1 more
TLDR
A combinatorial strategy to efficiently construct LVs using EGFP, hPlk2 wild type (WT) and mutant genes as inserts using BamH I site for the inserts and the amounts and ratios of the insert and vector DNA were optimized to increase monomeric ligation.Abstract:
Lentiviral vectors (LVs) are powerful tools for transgene expression in vivo and in vitro. However, the construction of LVs is of low efficiency, due to the large sizes and lack of proper clone sites. Therefore, it is critical to develop efficient strategies for cloning LVs. Here, we reported a combinatorial strategy to efficiently construct LVs using EGFP, hPlk2 wild type (WT) and mutant genes as inserts. Firstly, site-directed mutagenesis (SDM) was performed to create BamH I site for the inserts; secondly, pWPI LV was dephosphorylated after BamH I digestion; finally, the amounts and ratios of the insert and vector DNA were optimized to increase monomeric ligation. Our results showed that the total percentage of positive clones was approximately 48%±7.6%. Using this method, almost all the vectors could be constructed through two or three minipreps. Therefore, our study provided an efficient method for constructing large-size vectors.read more
Citations
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Intercalation and delamination of layered carbides and carbonitrides
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TL;DR: Intercalation of hydrazine, and its co-intercalation with N,N-dimethylformamide, resulted in increases of the c-lattice parameters of surface functionalized f-Ti3C2, from 19.5 to 25.48 and 26.8 Å, respectively.
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MAPbI3-xClx Mixed Halide Perovskite for Hybrid Solar Cells: The Role of Chloride as Dopant on the Transport and Structural Properties
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TL;DR: In this paper, the structural properties of iodide/chloride mixed-halide perovskites and correlating them with their photovoltaic performances were investigated, and they found that, independent of the components ratio in the precursor solution, Cl incorporation in an iodide-based structure, is possible only at relatively low concentration levels (below 3-4%).
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References
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Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis.
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Journal ArticleDOI
Germline Transmission and Tissue-Specific Expression of Transgenes Delivered by Lentiviral Vectors
TL;DR: Transgenic mice carrying the green fluorescent protein (GFP) gene driven by a ubiquitously expressing promoter are generated and transgenic rats that express GFP at high levels are generated, suggesting that this technique can be used to produce other transgenic animal species.