Control of triglyceride storage by a WD40/TPR-domain protein.

Obesity is a metabolic disorder related to improper control of energy uptake and expenditure, which results in excessive accumulation of body fat. Initial insights into the genetic pathways that regulate energy metabolism have been provided by a discrete number of obesity-related genes that have been identified in mammals. Here, we report the identification of the adipose (adp) gene, the mutation of which causes obesity in Drosophila.

Genetic analysis of Hispanic individuals with cystic fibrosis.

We have performed molecular genetic analyses of Hispanic individuals with cystic fibrosis (CF) in the southwestern United States. Of 129 CF chromosomes analyzed, only 46% (59/129) carry delta F508. The G542X mutation was found on 5% (7/129) of CF chromosomes.

Mutation analysis of the cystic fibrosis transmembrane regulator gene in Native American populations of the southwest.

We report DNA and clinical analyses of cystic fibrosis (CF) in two previously unstudied, genetically isolated populations: Pueblo and Navajo Native Americans. Direct mutation analysis of six mutations of the CFTR gene--namely, delta F508, G542X, G551D, R553X, N1303K, and W1282X--was performed on PCR-amplified genomic DNA extracted from blood samples. Haplotype analyses with marker/enzyme pairs XV2c/TaqI and KM19/PstI were performed as well.

Molecular analysis of cis-regulatory sequences at the alpha-amylase locus in Drosophila melanogaster.

The Amylase locus in Drosophila melanogaster contains duplicate, divergently transcribed structural genes for alpha-amylase, AmyA and AmyB. A sensitive and reliable transient expression assay was developed for testing amylase activities produced by exogenous Amy genes in somatically transformed larvae of an amylase-null strain of flies. Alleles tested, AmyA and AmyB, came from recombinant clone lambda Dm65, which contains genomic DNA from a Canton-S strain.

Tissue-specific and dietary control of alpha-amylase gene expression in the adult midgut of Drosophila melanogaster.

The regulatory effects of allelic substitution at the trans-acting mapP locus and of dietary glucose on the expression of the duplicate genes for alpha-amylase (Amy) in Drosophila melanogaster were examined in the anterior midgut and posterior midgut regions of mature flies. The levels of amylase activity and amylase protein, as well as the abundance of amylase-specific RNA, were quantified. All 3 parameters of Amy expression were concordant.

Dosage compensation and dietary glucose repression of larval amylase activity in Drosophila miranda.

The functional locus for alpha-amylase (Amy) in Drosophila miranda is in the evolutionarily new X2 chromosome. X2 evolved from an autosome in response to an ancestral autosome-Y translocation that gave rise to the "neo-Y" chromosome of this species. Y-linked Amy, if still present in the ancestrally translocated element, is unexpressed.

Amylase gene expression in intraspecific and interspecific somatic transformants of Drosophila.

The Amylase locus in Drosophila melanogaster normally contains two copies of the structural gene for alpha-amylase, a centromere-proximal copy, Amy-p, and a distal copy, Amy-d. Products of the two genes may display discrete electrophoretic mobilities, but many strains known to carry the Amy duplication are characterized by a single amylase electromorph, e.g.

Molecular genetics of a three-gene cluster in the Amy region of Drosophila.

Analysis of amylase RNA levels in the anterior and posterior midgut regions of flies from the Amy1,6 mapA and c Amy2,3 mapC strains of D. melanogaster, reared on yeast and on yeast supplemented with glucose, indicates that the trans-acting map gene controls the abundance of amylase RNA tissue-specifically, i.e.

Molecular cloning of alpha-amylase genes from Drosophila melanogaster. III. An inversion at the Amy locus in an amylase-null strain.

Overlapping clones of the structural gene region for alpha-amylase, Amy, were isolated from a lambda EMBL4 library containing genomic DNA fragments from an amylase-null strain of Drosophila melanogaster. Southern blot analysis and restriction endonuclease mapping of the cloned region indicate that it contains an Amy gene duplication within an inverted repeat sequence as is characteristic of the genomic arrangement for this species. Spacing between the cloned gene copies is similar to that commonly found in other strains.

Structural organization of the alpha-amylase gene locus in Drosophila melanogaster and Drosophila miranda.

Chromosomal sites belonging to the alpha-amylase gene family have been identified in D. melanogaster and D. miranda and in the sibling species of miranda, pseudoobscura, and persimilis.

Developmental analysis of lipids from wild-type and adipose60 mutants of Drosophila melanogaster.

A comparative developmental analysis was made of lipids from wild-type and adipose60 (adp60) mutants of Drosophila melanogaster. The lipid content and fatty acid profiles of late third instar larvae, pupae, and mature adults were characterized in methanol:chloroform extracts utilizing thin layer and gas-liquid chromatography. Total lipid content of mutant adults was approximately twice that of the wild-type, but no genotypic differences in lipid content were seen in earlier developmental stages.

Structural organization of the Amy locus in seven strains of Drosophila melanogaster.

Restriction maps were made by Southern blot analysis of the Amy (alpha-amylase) region in 7 strains of D. melanogaster using endonucleases SalI, XhoI and EcoRI. These were compared to the map of lambda Dm65 which contains the cloned Amy region.

Molecular cloning of alpha-amylase genes from Drosophila melanogaster. I. Clone isolation by use of a mouse probe.

A cloned alpha-amylase cDNA sequence from the mouse is homologous to a small set of DNA sequences from Drosophila melanogaster under appropriate conditions of hybridization. A number of recombinant lambda phage that carry homologous Drosophila genomic DNA sequences were isolated using the mouse clone as a hybridization probe. Putative amylase clones hybridized in situ to one or the other of two distinct sites in polytene chromosome 2R and were assigned to one of two classes, A and B.

Molecular cloning of alpha-amylase genes from Drosophila melanogaster. II. Clone organization and verification.

Restriction maps of an alpha-amylase structural gene clone, lambda Dm65, and of four putative alpha-amylase pseudogene clones are presented. Two alpha-amylase structural genes, inverted with respect to each other, are contained in lambda Dm65. Subregions of internal DNA sequence homology within lambda Dm65 and of cross-homology between the presumptive pseudogene clones and lambda Dm65 were determined.

Modification of wheat straw in a high-shear mixer.

Wheat straw (Ws)was treated in a pilot-scale continuous mixer to disrupt the lignin-hemicellulose-cellulose (LHC) complex. An efficient and practical method was desired to remove lignin and hemicellulose (pentosans)rapidly and efficiently from the lignocellulose complex and to make the cellulose accessible to enzymatic hydrolysis. Milled WS in the presence of various chemicals in aqueous solutions was extruded from the mixer under several processing conditions.

Hydrolysis of wheat straw hemicellulose with trifluoroacetic acid. Fermentation of xylose with Pachysolen tannophilus.

Treatment of wheat straw with 1N trifluoroacetic acid (TFA) for 7 h at reflux temperature yielded 23% xylose based upon initial straw weight. This corresponds to about an 80% xylose yield based on the xylan content of the hemicellulose. The cellulose component of wheat straw was largely unaffected, as evidenced by low glucose yields.

Liquid-phase dehydration of aqueous ethanol-gasoline mixtures.

Two-phase mixtures of gasoline, water, and ethanol were dehydrated with both starch and saponified starch-g-polyacrylonitrile (HSPAN). Whereas starch absorbed ethanol as well as water, HSPAN selectively absorbed the water component, allowing ethanol to dissolve in the gasoline phase.