遗传学实验技术

Expressed sequence tag (EST) surveys are an efficient way to characterize large numbers of genes from an organism. The rate of gene discovery in an EST survey depends on the degree of redundancy of the cDNA libraries from which sequences are obtained. We consider statistics for the comparison of EST ...

Expressed sequence tags (ESTs) are fragments of mRNA sequences derived through single sequencing reactions performed on randomly selected clones from cDNA libraries. To date, over 45 million ESTs have been generated from over 1400 different species of eukaryotes. For the most part, EST p ...

Expressed Sequence Tags (ESTs) are fragments of cDNA clones. They correspond to the most abundant type of cDNA information available in the public databases. ESTs have been used for expression profiling, gene identification, characterization of differentially expressed genes, and ...

Complementary DNA (cDNA) sequencing can be used to sample an organism’s transcriptome, and the generated EST sequences can be used for a variety of purposes. They are especially important for enhancing the utility of a genome sequence or for providing a gene catalog for a genome that has not or will not be ...

Sequencing of cDNA libraries is an efficient and inexpensive approach to analyze the protein-coding portion of a genome. It is frequently used for surveying the genomes of poorly studied eukaryotes, and is particularly useful for species that are not easily amenable to genome sequencing, b ...

Full-length cDNAs are essential for the correct annotation of transcriptional units and gene products from genomic sequence data and for functional analysis of the genes. Full-length cDNA libraries are very important resources for isolation of the full-length cDNAs. The biotinyla ...

Critical steps in a cDNA library preparation include efficient cDNA synthesis, selection of full-length cDNAs, normalizing their abundance, and the subtraction of redundant transcripts. The use of trehalose and sorbiol stabilizes the activity of the reverse transcriptase lead ...

A common task in EST projects is the conversion of sequence chromatograms originating from gel-based or capillary sequencers into annotated sequence objects. Here we describe the usage of a software pipeline (available from http://www.nematodes.org/bioinformatics/), which has ...

Expressed sequence tags (ESTs) have proven to be one of the most rapid and cost-effective routes to gene discovery for eukaryotic genomes. Furthermore, their multipurpose uses, such as in probe design for microarrays, determining alternative splicing, verifying open reading frames, a ...

Eukaryotes using trans-splicing for transcript processing incorporate a taxon-specific sequence tag (the spliced leader, SL) to a proportion (either all or a fraction) of their mRNAs. This feature may be exploited for the preparation of full-length-enriched cDNA libraries from these ...

The internal transcribed spacer (ITS) is the locus of choice with which to characterize fungal diversity in environmental samples. However, methods to analyze ITS datasets have lagged behind the capacity to generate large amounts of sequence information. Here, we describe our bioinfo ...

Affinity purification and mass spectrometry analyses have been used in various organisms to identify �protein complexes and determine protein–protein interactions in vivo. In comparison with the TAP (tandem affinity purification) tag, the 3� FLAG is a relatively small epitope tag. It ...

Deep transcriptome profiling of pathogen-infected tissues enhances the understanding of molecular mechanisms underlying host–pathogen interactions. Illumina’s next generation sequencing technology sequencing-by-synthesis (SBS) is a powerful tool to rapidly se ...

The model filamentous fungus Neurospora crassa has been the focus of functional genomics studies for the past several years. A high-throughput gene knockout procedure has been developed and used to generate mutants for more than two-thirds of the ∼10,000 annotated N. crassa genes. Yeast rec ...

For functional characterization of predicted genes encoding hypothetical proteins in fungal genomes, it is complementary to genetic studies to determine their expression and subcellular localization patterns in different developmental or infection stages. It is also impo ...

Phenotypic analysis of Neurospora crassa knockout (KO) mutants was used as a vehicle to introduce students to laboratory research. The availability of gene deletion strains was the impetus for the development of a program designed to introduce beginning science students to basic micro ...

With genome sequences of more and more fungi become available, high-throughput systematic �mutagenesis is desirable for functional genomics studies. While a number of random insertional mutagenesis and targeted gene disruption approaches have been used in filamentous fungi, A ...

Fusarium verticillioides is a major pathogen of corn and poses a significant risk to human health by producing mycotoxins that accumulate in kernels. Considerable efforts have focused on identifying genes involved in secondary metabolism and pathogenesis. The availability of a seq ...

The interaction of the filamentous fungal plant pathogen Cryphonectria parasitica with its virulence-attenuating viruses provides a unique platform to explore the molecular biology and genetics of virus–host interactions. Following the development of transformation p ...

Given recent technological advances, we are in a golden era of cell and whole organism research. With the availability of so many sequenced genomes, and the data that has been mined there-in, it is easy to gain the impression that all our work as scientists is complete. Instead, such work and results have now ...