Southern analysis of BT-R 1, the Manduca sexta gene encoding the receptor for the Cry1Ab toxin of Bacillus thuringiensis

Various subspecies of the gram-positive bacterium Bacillus thuringiensis are known to produce a wide array of insecticidal crystal proteins (ICPs) upon sporulation. These ICPs act primarily on the brush border of midgut epithelial cells of susceptible larvae. Recently, a protein of 210?kDa, isolated from the midgut of Manduca sexta, has been demonstrated to bind the Cry1Ab toxin produced by B. thuringiensis subsp. berliner and is therefore postulated to be involved in mediating the toxicity of Cry1Ab. The cDNA encoding the 210?kDa protein, termed BT-R₁ (Bacillus thuringiensis receptor-1), was recently cloned, and shows limited homology to the cadherin superfamily of proteins. Quite naturally, there is a great deal of interest in the characterization of BT-R ₁ , the gene encoding the 210?kDa Cry1Ab binding protein. The studies presented here involve the use of various restriction fragments prepared from the cDNA encoding BT-R₁ as probes of Southern blots bearing M. sexta genomic DNA cleaved with a variety of restriction endonucleases. These Southern blot data reveal that there are two discrete regions within the M. sexta genome which encode sequences homologous to BT-R₁. On the basis of the signal intensities seen on Southern blots, it appears that only one of these genes encodes BT-R₁, whereas the other is a closely related homologue.     

A [2Fe-2S] protein from the hyperthermophilic bacterium Aquifex aeolicus.

Overexpression in Escherichia coli of the fdx4 gene from Aquifex aeolicus has allowed isolation and characterization of the first hyperthermophilic [2Fe-2S](Scys)(4) protein, a homodimer of M = 2 x 12.4 kDa with one [2Fe-2S] cluster per subunit. This protein is undamaged by heating to 100 degrees C for at least three hours. The primary structure, in particular the characteristic distribution of the four cysteine ligands of the metal site, and the spectroscopic properties of the A. aeolicus protein relate it to well characterized [2Fe-2S] proteins from Clostridium pasteurianum and Azotobacter vinelandii. These proteins are also homologous to subunits or domains of hydrogenases and NADH-ubiquinone oxidoreductase (Complex I) of respiratory chains. The A. aeolicus [2Fe-2S] protein is thus representative of a presumably novel protein fold involved in a variety of functions in very diverse cellular backgrounds.     

Number and pattern of association of chromonemata in the chromosomes of Tradescantia

Summary Analysis of reconstructions, prepared from electron micrographs of successive longitudinal serial sections, has led to the conclusion that the somatic telophase chromosome of Tradescantia paludosa contains four cytologically separable chromonemata. The four represent a pair of pairs, that is, two diplospiremes — one with its two chromonemata arranged helically in dextrorse relationship, and the other with its two in sinistrorse relationship — which are associated to form a tetraspireme. During anaphase and telophase the tetraspireme constitutes the chromosome; during prophase and metaphase the tetraspireme represents one of the two chromatids of the chromosome, which is accordingly an octospireme in terms of the number of cytologically identifiable chromonemata. Loose intertwining of the two tetraspiremes during late prophase accounts for the so-called relational coiling.This paper is dedicated to Professor Hans Bauer on his sixtieth birthday anniversary in appreciation of his contributions to the development of modern cytology.The work reported here was supported in part by Research Grants GM-10499 from the National Institutes of Health, U.S. Public Health Service, and GB-290 from the National Science Foundation, and in part by a NATO fellowship awarded to E. Sparvoli by the Italian National Council of Research.