Screening for single nucleotide polymorphisms in highly degraded DNA by using the amplified fragment length polymorphism technique.

Short tandem repeat (STR) analysis is generally used for human identification of forensic samples; however, standard STR analysis sometimes fails to generate full profiles since DNA is frequently degraded by various environmental factors. Recently, single nucleotide polymorphism (SNP) analysis has attracted attention for human identification since the shorter amplicons are better suited for degraded samples. Though various SNP loci are used for analysis of degraded samples, it is unclear which ones are more appropriate.

Investigation of single nucleotide polymorphism loci susceptible to degradation by ultraviolet light.

DNA in biological fluids is often degraded by environmental factors. Given that single nucleotide polymorphism (SNP) analyses require shorter amplicons than short tandem repeat (STR) analyses do, their use in human identification using degraded samples has recently attracted attention. Although various SNP loci are used to analyze degraded samples, it is unclear which ones are more appropriate.

Characterization of cellular and extracellular DNA in saliva.

Although the presence of extracellular DNA in various body fluids was discovered long ago, only recently has it begun to attract attention for examining the genetic profiles of individuals in forensics studies. However, information on extracellular DNA is scarce. Among human body fluids, saliva is known to be rich in extracellular DNA.

A simple ABO genotyping by PCR using sequence-specific primers with mismatched nucleotides.

In forensics, the specific ABO blood group is often determined by analyzing the ABO gene. Among various methods used, PCR employing sequence-specific primers (PCR-SSP) is simpler than other methods for ABO typing. When performing the PCR-SSP, the pseudo-positive signals often lead to errors in ABO typing.

Changes in the brain after intracerebral implantation of a lead pellet in the rat.

In gunshot wounds to the head, the bullet containing neurotoxic lead may remain in the brain after trauma, and brain damage is therefore anticipated. We developed an animal model incorporating a lead ball implanted in the brain, or a glass ball as a control, and analyzed histological and biochemical changes in the brain for 28 days after surgery. The concentration of lead in the brain increased with time after implantation of the lead ball, while lead was not detected in brains implanted with a glass ball.

Selection of a biotin protein ligase by phage display using a combination of in vitro selection and in vivo enzymatic activity.

We report the development of a phage display-based method for the direct selection of enzymatic activity by a combination of selection in vitro and an assay of enzymatic activity in vivo. We describe the selection of a biotin protein ligase (BPL) that specifically recognizes neurotensin as a substrate, as an example of the utility of our method. We constructed an enzyme library with a diversity of 8.

Selection of an active enzyme by phage display on the basis of the enzyme's catalytic activity in vivo.

We have developed a novel phage display method based on catalytic activity for the in vivo selection of an enzyme. To confirm the validity of our method and to demonstrate its potential utility, we used biotin protein ligase (BPL) from Escherichia coli as a model enzyme. We were able to demonstrate the potential value of our method by selective enrichment for the birA gene, which encodes BPL, in a mixed library.

Reversed-phase HPLC determination of chlorophyll a' and phylloquinone in Photosystem I of oxygenic photosynthetic organisms. Universal existence of one chlorophyll a' molecule in Photosystem I.

Chlorophyll (Chl) a', the C132-epimer of Chl a, is a constituent of the primary electron donor (P700) of Photosystem (PS) I of a thermophilic cyanobacterium Synechococcus (Thermosynechococcus) elongatus, as was recently demonstrated by X-ray crystallography. To determine whether PS I of oxygenic photosynthetic organisms universally contains one molecule of Chl a', pigment compositions of thylakoid membranes and PS I complexes isolated from the cyanobacteria T. elongatus and Synechocystis sp.