Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:2.7.1.21 (thymidine kinase)
 7,561 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Targeted gene replacement via homologous recombination (HR) is a conventional approach for the analysis of gene function. However, this event is rare in Beauveria bassiana, which hampers efficient functional analysis in this widely used entomopathogenic fungus. To improve homologous recombination frequency in B. bassiana, we investigated the effect of the ratio of homologous sequence to non-homologous sequence (HS/NHS) in gene disruption cassette upon the HR frequency by two gene loci BbNtl and BbThi, using the herpes simplex virus thymidine kinase as a negative selectable marker against ectopic transformants. Our data revealed that an increase of the ratio of HS/NHS achieved by either extending homologous sequence or decreasing non-homologous sequence could improve HR frequency in B. bassiana. We determined empirically that (1) at least 700 bp of homology to both sides of a target gene was needed to get a reasonable number of disruptants, e.g., 6.7 per thousand to 13.3 per thousand in B. bassiana. (2) When the ratio of HS/NHS was above 0.8, an acceptable HR frequency could be achieved for gene replacement in B. bassiana, while when the ratio was below 0.3, few gene disrupted mutants were obtained.
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PMID:The size and ratio of homologous sequence to non-homologous sequence in gene disruption cassette influences the gene targeting efficiency in Beauveria bassiana. 1914 36

To be able to label a gene and monitor its migration are key important approaches for the clinical application of cancer suicide gene therapy. Photonic nanomaterials are introduced in this work. One of the most promised suicide genes - herpes simplex virus thymidine kinase (HSV-TK) gene - is successfully linked with CdTe/CdS core/shell quantum dots (QDs) via EDC/NHS coupling method. From confocal microscopy it was demonstrated that plasmid TK intracellular trafficking can be effectively and distinctly traced via monitoring the luminescence of the QDs up to 96 h after transfection of QDs-TK conjugates into Hela cells. MTT results show that the QDs-TK conjugates have a high efficient cytotoxicity after adding GCV into Hela cells, whereas the QDs exert no detectable deleterious effects on the cellular processes. The apoptosis induced by QDs-TK conjugates with GCV is distinctly traced partly due to the strong luminescence of the QDs. Our results indicate that photonic nanomaterials, e.g. QDs, provide a tool for monitoring TK gene delivery and anti-cancer activity.
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PMID:Monitoring HSV-TK/ganciclovir cancer suicide gene therapy using CdTe/CdS core/shell quantum dots. 2244 46