Improved Ligation Specificity with Chemically Modified Ligation Components
Poster Oct 04, 2012
Sabrina Shore, Alexandre Lebedev, Elena Hidalgo Ashrafi, Gerald Zon, Natasha Paul, Richard Hogrefe
Ligases are gaining utility in molecular biology applications, such as nucleotide sequence detection, single nucleotide polymorphism (SNP) detection, protein detection and “next generation” sequencing by ligation. With the increased demand for DNA ligases in the field of biotechnology, comes increased demand for ligation fidelity. Described approaches to improved ligation fidelity include ligases from different biological sources, point mutations of key amino acid residues within the ligase, modified reaction conditions and addition of crowding reagents, such as PEG. Although most approaches to improved ligation fidelity have focused on the ligase itself, further improvements are needed and may be attainable by a different approach. Herein a strategy to improve the discrimination between matched and mismatched targets is described which employs chemical modification to the nucleic acid components of the reaction, such as the donor probe, the acceptor probe and the ATP cofactor. The results demonstrate that chemically modified components increase the stringency of DNA ligase-mediated nucleic acid detection, providing a unique approach for SNP genotyping.
P450 Induction in Cryopreserved Hepatocytes from PXR and CAR Nuclear Receptor Knock-out RatsPoster
The nuclear receptors pregnane X receptor (PXR) and constitutive androstane receptor (CAR) are closely related transcription factors that regulate the expression of phase I (cytochrome P450s), phase II metabolizing enzymes and transporter genes in response to xenobiotics, including prescription drugs.READ MORE
Psychiatric Risk Gene Cacna1c and Early Life Stress: Potential Gene-Environment interactions?Poster
Early life stress (ELS) is highly associated with development of psychopathology
and mood disorders in adulthood. Genetic studies have identified variation in the gene calcium voltage-gated channel subunit alpha1C (CACNA1C) to increase risk for several psychiatric disorders. This poster assessed the expression of Cacna1c following prepubertal stress.
The Role of K13 in Artemisinin ResistancePoster
Plasmodium falciparum is evolving resistance to Artemisinin Combination Therapy. The gene with the strongest association with resistance is K13. K13 is an ortholog of the well characterized transcriptional regulator Keap1. In this work we transcriptionally characterized a mutant with a transposon inserted in the K13 promoter region which results in dysregulation of K13 at 2 points of the intraerythrocytic cycle of the life-cycle to identify the processes regulated by K13.READ MORE