Overexpression of bacterial RecA protein stimulates homologous recombination in somatic mammalian cells

The pairing of homologous molecules and strand exchange is a key event in homologous recombination promoted by RecA protein in Escherichia coli. Structural homologs of RecA are widely distributed in eukaryotes including mouse and man. As has been shown, human HsRad51 protein is not only structural but also functional homolog of RecA. The question arises whether the bacterial functional homolog of Rad51 can function in mammalian cells and increase the frequency of the homologous recombination. To investigate possible effects of bacterial RecA protein on the frequency of homologous recombination in mammalian cells, the E. coli RecA protein fused with a nuclear location signal from the large T antigen of simian virus 40 was overexpressed in the mouse F9 teratocarcinoma cells. We found that the frequency of gene targeting at the hprt locus was 10-fold increased in the mouse cells expressing the nucleus-targeted RecA protein. Southern blot analysis of individual clones that were generated by targeting recombination revealed predicted type of alterations in hprt gene. The data indicate that the bacterial nucleus-targeted RecA protein can stimulate homologous recombination in mammalian cells.     

Influence of Exon Duplication on Intron and Exon Phase Distribution

Nonrandomness in the intron and exon phase distributions in a sample of 305 human genes has been found and analyzed. It was shown that exon duplications had a significant effect on the exon phase nonrandomness. All of the nonrandomness is probably due to both the processes of exon duplication and shuffling. A quantitative estimation of exon duplications in the human genome and their influence on the intron and exon phase distributions has been analyzed. According to our estimation, the proportion of duplicated exons in the human genome constitutes at least 6% of the total. Generalizing the particular case of exon duplication to the more common event of exon shuffling, we modeled and analyzed the influence of exon shuffling on intron phase distribution. Received: 28 March 1997 / Accepted: 9 July 1997     

Physiological Heterothallism and Sexuality in Euascomycetes: A Partial History

Copyright 1998 Academic Press.     

The effect of gamma radiation on the enzyme activity of collalitin during storage

A study was made of the effect of gamma-irradiation of collalitin samples with doses of 25 and 50 kGy on proteolytic and enzyme activity with respect to denaturated collagen. The enzyme properties were found to change upon storage for periods of 6 and 12 months at 15 degrees C under unsterile conditions. No significant changes in the molecular mass of the collalitin samples were revealed by the method of gell-chromatography after irradiation and storage.     

Widespread positive selection in the photosynthetic Rubisco enzyme

Background  

Rubisco enzyme catalyzes the first step in net photosynthetic CO₂ assimilation and photorespiratory carbon oxidation and is responsible for almost all carbon fixation on Earth. The large subunit of Rubisco is encoded by the chloroplast rbcL gene, which is widely used for reconstruction of plant phylogenies due to its conservative nature. Plant systematicists have mainly used rbcL paying little attention to its function, and the question whether it evolves under Darwinian selection has received little attention. The purpose of our study was to evaluate how common is positive selection in Rubisco among the phototrophs and where in the Rubisco structure does positive selection occur.     

Gene flow and species cohesion following the spread of Schiedea globosa (Caryophyllaceae) across the Hawaiian Islands

Island radiations are often regarded as natural laboratories that allow us to study evolution in action. The genus Schiedea (Caryophyllaceae) is one of the largest radiations of angiosperms in the Hawaiian Islands, and Schiedea globosa is one of the few species in the genus to be found on more than one of the main islands. DNA sequences from nineteen nuclear and three chloroplast regions show a pattern of colonization from older to younger islands (west to east), with a concomitant decrease in genetic diversity eastwards (π=0.53% for O'ahu, 0.43% for Moloka'i and 0.36% for Maui). While polymorphisms in the maternally inherited chloroplast have become fixed on different islands (F(ST)=0.804), significant gene flow between islands is inferred for the nuclear genome (F(ST)=0.270). This gene flow appears to be uneven, with most gene flow outwards from the central island. The extent of inter-island gene flow through wind pollination was assessed in an isolation-migration framework; the inferred rate, c. 1 migrant per generation, may be sufficient to prevent divergence of S. globosa populations and ensure cohesion of the species following the colonization of new islands.     

Pulse-electrotypes of Listeria monocytogenes strains, isolated in Moscow

The characterization of the pulse-electrotypes of L. monocytogenes, isolated in 2003-2004 in Moscow from different sources, is presented. Among the cultures, isolated from humans, one outbreak pulse electrotype was detected and from different objects in buildings where a wide variety of food products was produced several probably related and unrelated pulse-electrotypes were obtained. The conclusion was made that several independent L. monocytogenes clones existed on the territory of Moscow, and many products supplied to retail trade and public catering enterprises were contaminated with these clones. Pulse electrophoresis was shown to be the most effective method for intraspecific typing and the study of the molecular epidemiology of listeriosis. Grounds for the necessity to improve the microbiological diagnostics of L. monocytogenes infection are given.     

Characterization of Escherichia coli strains O157:H7, isolated on the territories of the Central Federal District

The characterization of E.coli strains O157:H7, isolated from humans and animals on some territories of the Central Federal District, is presented. Among the isolates from human outbreaks, related and, probably, related cultures prevailed, while among the isolates obtained from different animals mainly unrelated cultures have been detected. A conclusion has been made concerning the existence of several independent zoonotic reservoirs of E. coli O157:H7 infection on this territory. The advantages and drawbacks of the use of pulse electrophoresis in the characterization of E. coli O157:H7 are discussed. Grounds are given for the necessity of the patients examination with hemorrhagic enetrocolitis for the presence of E. coli O157:H7, as well as for the expediency of having a special item for the registration of this E. coli infection in relevant statistical forms.     

DNA diversity in Hawaiian endemic plant Schiedea globosa

This is the first report of a study devoted to the population genetics of speciation in the endemic Hawaiian plant genus Schiedea (Caryophyllaceae). Here, we report the estimates of DNA sequence diversity and divergence in a newly isolated nuclear gene from Maui and Oahu Schiedea globosa populations. Overall, the species-wide average heterozygosity per silent site is pi = 0.3%. The silent DNA diversity on the older island of Oahu (pi = 0.24%) is almost twice as high as on the younger Maui (pi = 0.14%). Consistent with this, the haplotype phylogeny suggests a more recent origin of the Maui populations. There is no significant isolation between the two Maui populations (F(st)=0.027), while isolation between the two islands is high (F(st)=0.57, P<0.0001). Pairwise mismatch distributions suggest population growth approximately 660 and 310 thousand generations ago for the Oahu and the Maui populations, respectively, which may be the minimal age for these populations. This is consistent with a fairly neutral frequency spectrum (Tajima's D is 0.34 and -0.94 for the Oahu and the Maui populations, respectively), suggesting that both populations are sufficiently old to have recovered from any initial founder effects. Relatively high nuclear DNA diversity in the S. globosa populations illustrates the usefulness of a DNA sequence-based approach to the population genetics of island plant populations.