Human XPA and XRCC1 DNA repair proteins expressed in yeast, Saccharomyces cerevisiae.

Human XPA and XRCC1 DNA repair proteins have been expressed in a series of novel yeast episomal vectors. Expression of XPA cDNA resulted in synthesis of anti-XPA crossreacting polypeptides of 40 and 42 kDa, the status of the native protein found in human cells. Likewise, the majority of the recombinant XRCC1 found in the yeast intracellular fraction corresponded to the molecular mass of the full-length human protein.

An easy and accurate agarose gel assay for quantitation of bacterial plasmid copy numbers.

An assay for quantitation of plasmid copy numbers in bacterial cell cultures has been developed and validated. The method combines isolation of total bacterial DNA (including both plasmid and genomic DNA), running a series of twofold dilutions of total DNA in an agarose gel followed by ethidium bromide staining, and subsequent scanning of the gel picture negatives. We have developed a novel set of rules for integration of the scan data that allows us to achieve high assay precision, accuracy, and sensitivity.

Quantitative restriction fragment length polymorphism: a procedure for quantitation of diphtheria toxin gene CRM197 allele.

Here we present an assay for quantitation of a particular gene allele in DNA mixtures by means of restriction fragment length polymorphism (RFLP) in combination with polymerase chain reaction (PCR). We applied the quantitative RFLP principle for estimation of the relative amount of diphtheria toxin gene CRM197 allele in Corynebacterium diphtheriae culture DNA samples. The procedure is based on PCR-mediated generation of an artificial AluI restriction site specifically with the CRM197 DNA template.

TAP2 alleles in inflammatory arthritis.

Sixty patients with reactive arthritis (ReA) and 40 with rheumatoid arthritis (RA), were typed for H LA-B27 and class II antigens DR and DQ, and studied for TAP2 gene polymorphism in comparison with 60 healthy controls. TAP2 polymorphisms at positions 379, 565, 665, and 687 were analyzed using amplification refractory system-based PCR and polymorphisms at positions 386 and 651 using oligonucleotide hybridization. The frequency of the TAP2A/A genotype was 30%(12/40) in RA, in contrast to 13% (8/60) in the controls.

CYP2D6 genotyping in a Russian population using a novel approach for identification of the CYP2D6A mutation.

The frequency of 29A and 29B mutations in the CYP2D6 gene, the most common mutations among Caucasoid PM (debrisoquine 4-hydroxylase defficient) individuals, has been analyzed in the Russian population. For the detection of the 29A mutation, a new one-step ARMS PCR approach has been developed. The frequency of the 29B mutant allele in Russians appeared to be significantly lower than in other Caucasoid populations, and this observation can be explained by the mixed origin of the population inhabiting Russia.

Frequency of a specific cytochrome P4502D6B (CYP2D6B) mutant allele in clinically differentiated groups of patients with Parkinson disease.

The frequency of the cytochrome P4502D6B CYP2D6B (29B) mutant allele has been determined in three clinically distinct groups of patients with Parkinson disease. No differences in mutation frequency among the patients with the rigidity-akinetic and monosymptomatic tremor forms has been observed compared to the healthy control group, while in the group with akinetic-rigidity tremor symptoms the frequency of heterozygous wt/29B individuals was significantly increased. Therefore, individuals bearing the CYP2D6B mutation appear to be predisposed to the development of this particular form of Parkinson disease, and the presence of the 29B mutation in the genotype may serve as an additional diagnostic criteriaum for the clinical differentiation of patients with Parkinson disease.

A rapid and simple DNA fingerprinting method using RFLP and SSCP analysis of the hypervariable noncoding region of human mitochondrial DNA.

A simple method for identification of individuals and maternity testing has been developed. This technique includes PCR amplification of the 199-bp hypervariable portion of the noncoding region of human mtDNA, digestion with RsaI and subsequent SSCP analysis of restriction fragments with DNA silver staining. Using this approach we have analysed the DNA fingerprint patterns of the family members.

Comparative analysis of spontaneous mitotic recombination in [cir0] and [cir+] strains of the yeast Saccharomyces cerevisiae.

The influence of the 2 microns plasmid on homologous recombination in the right arm of chromosome XV of the yeast Saccharomyces cerevisiae has been examined. No differences between spontaneous mitotic recombination rates in [cir0] and [cir+] derivatives of two yeast diploid tester strains were detected. In the course of analysis an unusually high coincident conversion frequency at ADE2, HIS3, and two RFLP loci adjacent to ADE2, was observed.