The primary structure of cutinase, an extracellular fungal enzyme involved in the penetration of plants by pathogenic fungi, has been determined from the nucleotide sequence of cloned cDNA. Clones containing cDNA made from poly(A)(+) RNA isolated from fungal cultures induced to synthesize cutinase were screened for their ability to hybridize with the [(32)P]cDNA for mRNA unique to the induced c...

Determining the number of fluorescent entities that are coupled to a given molecule (DNA, protein, etc.) is a key point of numerous biological studies, especially those based on a single molecule approach. Reliable methods are important, in this context, not only to characterize the labeling process but also to quantify interactions, for instance within molecular complexes. We combined fluoresc...

Name of the Sequence. Stearoyl coenzyme A desaturase (SCD) cDNA. Genus and Species. Capra hircus. Origin of Clone. Total RNA was isolated from a mammary gland sample from a lactating Murciano-Granadina goat using Trizol reagent (Life Technologies, Rockville, MD) and following supplier’s instructions. The first strand of the cDNA was synthetized using the M-MuLV reverse transcriptase (Life Techn...

To examine the control of porcine von Willebrand factor (vWF) biosynthesis we cloned human vWF complementary DNA (cDNA) and investigated the expression of the vWF gene in lungs from normal pigs and pigs with severe von Willebrand's disease (vWD). Recombinant clones spanning approximately 90% of human vWF cDNA were identified in a lambda gt10 human lung cDNA library by screening with oligonucleo...

MAS (marker assisted selection) is useful for cultivar development in the lower plant Saccharina japonica . In this study, 167 screened SSR markers and 125 individuals were used to construct a S.japonica genetic linkage map with 27 groups total Haldane length of 2069 cM. Constructed aligned assembled pseudo-chromosomes S. genome sequences (NCBI: MEHQ00000000) anchored 23 31 pseudo-chromosomes. ...

Lysophosphatidic acid acyltransferase acylates the sn-2 hydroxyl group of lysophosphatidic acid to form phosphatidic acid, a precursor to triacylglycerol. A cDNA encoding lysophosphatidic acid acyltransferase was isolated from developing seeds of meadowfoam (Limnanthes alba alba). The cDNA encodes a 281-amino acid protein with a molecular mass of 32 kD. The cDNA was expressed in developing seed...

The genetic defects in Prader-Willi syndrome (PWS) and Angelman syndrome (AS) map to 15q11-13. Using microdissection, we have recently isolated several DNA probes for the critical region. Here we report that microclone MN7 detects multiple loci in 15q11-13 and 16p11.2. Eight yeast artificial chromosome (YAC) clones, two genomic phage clones, and two placenta cDNA clones were isolated to analyze...

Material and methods For this purpose, we adopted the strategy of RT-PCR using degenerated primers obtained from sequences of known plant OSC genes. Total RNA was extracted from leaves of M. ilicifolia and used in the synthesis of cDNA. The PCR amplification of core fragments of OSC genes was performed with partially degenerated primers designed to anneal to highly conserved regions among OSC g...

We have developed a new strategy for producing subtracted cDNA libraries that is optimized for connective and epithelial tissues, where a few exceptionally abundant (super-prevalent) RNA species account for a large fraction of the total mRNA mass. Our method consists of a two-step subtraction of the most abundant mRNAs: the first step involves a novel use of oligo-directed RNase H digestion to ...

We describe the cloning of double-stranded cDNA synthesized from lactating human mammary gland total poly(A)-containing RNA, into the EcoRI site of the plasmid pAT153. Nine recombinants were shown to contain alpha-lactalbumin cDNA sequences as determined by positive hybridisation translation of complementary RNA. Restriction enzyme maps were determined for six of these. Alignment of the restric...