Trichoderma reesei sequences that bind to the nuclear matrix enhance transformation frequency

Three DNA fragments, trs1, 2 and 3, were isolated from the Trichoderma reesei genome on the basis of their ability to promote autonomous replication of plasmids in Saccharomyces cerevisiae. Each trs element bound specifically to the isolated T. reesei nuclear matrix in vitro, and two of them bound in vivo, indicating that they are matrix attachment regions (MARs). A similar sequence previously isolated from Aspergillus nidulans (ans1) was also shown to bind specifically to the T. reesei nuclear matrix in vitro. The T. reesei MARs are AT-rich sequences containing 70%, 86% and 73% A+T over 2.9, 0.8 and 3.7 kb, respectively for trs1, 2 and 3. They exhibited no significant sequence homology, but were shown to contain a number of sequence motifs that occur frequently in many MARs identified in other eukaryotes. However, these motifs occurred as frequently in the trs elements as in randomly generated sequences with the same A+T content. trs1 and 3 were shown to be present as single copies in the T. reesei genome. The presence of the trs elements in transforming plasmids enhanced the frequency of integrative transformation of T. reesei up to five fold over plasmids without a trs. No evidence was obtained to suggest that the trs elements promoted efficient replication of plasmids in T. reseei. A mechanism for the enhancement of transformation frequency by the trs elements is proposed. Received: 1 March 1997 / Accepted: 13 May 1997     

High performance liquid chromatography,chemical laboratory practice: By Heinz Engelhardt. Springer-Verlag,New York/Berlin, 1979. 248 pp. $29.80

A gel filtration method has been developed for the complete removal of sodium dodecyl sulfate (SDS) from proteins and peptides. The protein or peptide (20 μg–10 mg) containing SDS (up to 30–60 mg) is dissolved in a mixture of propionic acid, formic acid, and water (2:1:2, $\text{v}\text{v}$). Under these conditions, protein-SDS (or peptide-SDS) complexes, as well as SDS micelles, are dissociated. Subsequently, protein and SDS can be separated on a small Sephadex G-25 superfine column. The recovery of protein is typically 90% or more.     

Improved assay of coenzyme F420 analogues from methanogenic bacteria

Summary An improved method for separating analogues of coenzyme F₄₂₀ by isocratic reversed-phase high performance liquid chromatography is described. The method offers improved resolution, shorter chromatography runs and requires less complex apparatus.     

Genetic Variation and the Reproduction of Cordylanthus maritimus ssp. maritimus to Sweetwater Marsh, California

This study evaluated the genetic consequences of a reintroduction of the endangered annual plant Cordylanthus maritimus ssp. maritimus to Sweetwater Marsh (San Diego County, California). A survey of 21 enzyme loci in natural populations revealed that genetic diversity is very low and is primarily found as rare alleles at a few loci, making this species especially susceptible to the loss of alleles and heterozygosity through genetic drift. The reintroduction was performed in 1991 and 1992 by sowing seeds (collected from Tijuana Estuary) in numerous small patches of suitable habitat. For this study, leaf tissue was collected from all plants in all patches during flowering in 1995 and surveyed for genotype at the three enzyme loci that are polymorphic at Tijuana Estuary. Rare alleles were absent in 27 out of 30 patches for Pgm-1, in 17 out of 30 patches for Pgm-2, and in 10 out of 11 patches for Mdh-1. In all, half of the patches lacked any rare allele. Rare alleles tended to occur in patches with few individuals. Overall rare allele frequency was lower than in the colonies from which seeds were collected at two of the three loci, and heterozygosity was reduced. The Sweetwater Marsh population is at risk of losing most of its genetic variation at enzyme loci through the extinction of patches with few individuals. Future reintroduction attempts should attempt to create contiguous sets of patches or to periodically reseed existing patches to reduce the loss of genetic variation.     

Accelerated disulfide reduction with polyunsaturated fatty acids: a mechanism of ionic channel modulation?

Summary

Polyunsaturated fatty acids (PUFAs) have been shown to modulate the activity of ionic channels by an unknown mechanism. Some channels are activated (i.e. certain delayed-rectifier, potassium channels) and others are inhibited (i.e. certain calcium, sodium and other potassium channels). We have previously demonstrated that PUFAs can act as electron carriers. It is known that ionic channels can be redox modulated. The ability of fatty acids to serve as electron shuttling agents is proportional to their unsaturation. These PUFAs cause reduction of disulfides through a superoxide radical-independent mechanism, probably related to enhanced electron delocalization. The present study shows that there is a strong correlation between the ability of a PUFA to transfer an electron to a disulfide and its reported ability to modulate ionic channels. This suggests that electron transfer could be the mechanism of PUFAs action on particular ionic channels.     

Blastomycosis in wild wolves

Blastomycosis was fatal to a wild wolf in Minnesota, and serologic evidence of blastomycosis was found in a Wisconsin wolf. No unusual movements were detected in the Minnesota animal from October 1983 through October 1985. However, by early December 1985, this wolf was weak and debilitated, and it perished on 14 December after approaching a human residence.