Strict co-linearity of genetic and protein folding domains in an intragenically duplicated rat lens γ-crystallin gene

Two complementary DNA clones pRLγ-2 and pRLγ-3 of different rat lens γ-crystallin messenger RNAs have been used to identify γ-crystallin gene sequences in rat genomic DNA. Subsequently, the DNA present in the 18,000 to 20,000 bases region of the EcoRI digest, giving rise to a strong doublet hybridization signal, was cloned in λ phage Charon-4A. One of the clones, λRCHγ-3, carrying an insert of 17,500 bases has been characterized in detail. From analysis at the restriction enzyme level with 5′-, “middle” and 3′-specific subprobes of pRLγ-3 it could be deduced that λRCHγ-3 contains only one γ-crystallin gene. The coding sequences of this gene are interrupted by intronic DNA. The primary structure of this gene and its flanking regions have been established by sequencing the relevant regions of a subclone of λRCHγ-3, designated pRCHγ-3.1. The sequence data show that the γ-crystallin gene extends over 2700 bases of rat genomic DNA. The gene is split by two introns, one of 87 base-pairs after the third translation codon and a large one of 1880 base-pairs after codon 84. The mosaic structure of the gene is strictly co-linear with the structure of the γ-crystallin polypeptide in that the large intron is positioned in a region which specifies the so-called “connecting peptide” and which links the two highly symmetrical and homologous protein domains. Although expected from the cDNA and protein sequence no introns were observed between the coding regions in the DNA specifying the two homologous folding motifs present in each protein domain. The relevance of this phenomenon in terms of the evolution of the mature γ-crystallin gene is discussed.     

Developmental biochemistry of cottonseed embryogenesis and germination

Plant Molecular Biology - DNAs complementary to the mRNAs coding for the major cotton 48 kD and 52 kD storage proteins have been cloned and used to characterize the principal cotton storage protein...     

Temperature regulation and water metabolism in the elephant shrew Elephantulus edwardi

1. The Cape elephant shrew Elephantulus edwardi maintained a stable body temperature (37.6 C) over a wide range of ambient temperatures. 2. Normal eutherian mechanisms of temperature regulation were employed in the maintenance of homeothermy. 3. Water turnover rate and metabolic rate were lower than predicted for a similar-sized eutherian. 4. Maximal urine concentration was fairly high (3118 +/- 267 mOsm/kg) with urine osmolality largely dependent on the urea concentration. 5. Relative medullary thickness of the kidney (6.61 +/- 0.84) indicated a higher maximum urine concentration than found in the laboratory and reflected an arid habitat. 6. E. edwardi can be classed among the "advanced" homeotherms in view of the physiological adaptations employed for survival.     

The Avoidance Response in Phycomyces

Phycomyces sporangiophores grow away from stationary objects, a phenomenon known as the avoidance response. Evidence is presented suggesting that a growth-stimulating gas is emitted from the sporangiophore and is then swept to the leeward side by air currents resulting in higher gas concentration on that side. The presence of a stationary barrier decreases the passive movement of the gas away from the leeward side. It is proposed that an increase of this gas on one side causes that side to grow faster. Indirect evidence suggests that the gas is water vapor.     

Dissociation by Chelating Agents and Substructure of the Thermophilic Bacteriophage TP84

The thermophilic bacteriophage TP84 is dissociated into its head, tail, and released deoxyribonucleic acid (DNA) by chelating agents such as ethylenediaminetetraacetic acid (EDTA) and phosphate. The phage is more sensitive to EDTA than to phosphate, and dialysis against either agent causes more effective dissociation than standing in their presence. The tail possesses a knobbed structure which is inserted into the head of the intact phage and to which the DNA appears to be attached. The method of dissociating TP84 described in this paper provides a source of undamaged structural components and intact strands of DNA for subsequent investigations. A possible mechanism of chelate inactivation is discussed.     

Transformation in Bacillus amyloliquefaciens

The methodology and some of the requirements for the deoxyribonucleic acid-mediated transformation of an arginine auxotroph of Bacillus amyloliquefaciens to prototrophy are described.     

Chemically modified nylons as supports for enzyme immobilization. Polyisonitrile-nylon

Four-component condensations between amine, carboxyl, isocyanide and aldehyde lead to the formation of N-substituted amides (Ugi, 1962). The present paper describes the use of such condensations for the introduction of chemically reactive groups on to the polyamide backbone of nylon. Polyisonitrile-nylon was synthesized by partial hydrolysis of nylon-6 powder, followed by resealing of the newly formed -CO(2)... NH(2) (-) pairs via a four-component condensation, by using acetaldehyde and 1,6-di-isocyanohexane. Polyisonitrile-nylon could also be converted into a diazotizable arylamino derivative, polyaminoaryl-nylon, by a four-component condensation by using a bifunctional amine, pp'-diaminodiphenylmethane, in the presence of an aldehyde and a carboxylate compound. The versatility of four-component condensations involving the isocyanide functional group of polyisonitrile-nylon allowed coupling of proteins, in an aqueous medium at neutral pH, through either their amino or carboxyl groups. Trypsin and papain were bound to polyisonitrile-nylon through their amino groups by a four-component condensation by using acetaldehyde and acetate; conversely, succinyl-(3-carboxypropionyl-)trypsin, pepsin and papain were coupled through their carboxyl groups in the presence of acetaldehyde and an amine (Tris). Diazotized polyaminoaryl-nylon could be utilized for the immobilization of papain, via the tyrosine residues of the enzyme.     

Surface Structure of Bacillus stearothermophilus Ribosomes

No significant differences were noted in either the size or in the arrangement of the surface filaments of the ribosomes of Bacillus stearothermophilus and Escherichia coli.     

Electrophoretic and Immunological Differences Between the Cytochrome c3 of Desulfovibrio desulfuricans and that of Desulfovibrio vulgaris

The cytochrome c(3) of Desulfovibrio desulfuricans and that of D. vulgaris were purified to homogeneity as judged by disc gel electrophoresis and by ultracentrifugation. Both cytochromes had an oxidation-reduction potential of -205 +/- 5 mv at pH 7.0 and showed characteristic absorption bands at 525 and 553 nm in the reduced state. The molecular weights of the two cytochromes (calculated from sedimentation and diffusion data) were similar, with values of 13,500 to 14,300 for D. desulfuricans and 13,800 to 14,700 for D. vulgaris. The two cytochromes differed in their electrophoretic properties on Geon and polyacrylamide gel electrophoresis and did not share a common precipitating antigenic determinant as judged by immunodiffusion data.