遗传学实验技术

Telomeres are the sequences that form the ends of eukaryotic chromosomes and are essential structures that confer genome stability and guide chromosome behavior. In addition, the terminal regions of the chromosomes tend to house genes with predicted roles in ecological adaptation. U ...

Decoding the genome sequence is becoming a fundamental tool for molecular, genetic, and genomic studies. This chapter reviews the history of DNA sequencing and technical principles of different sequencing platforms, and compares the strengths and weaknesses of different techniq ...

The continued fast pace of fungal genome sequence generation has enabled proteomic analysis of a wide variety of organisms that span the breadth of the Kingdom Fungi. There is some phylogenetic bias to the current catalog of fungi with reasonable DNA sequence databases (genomic or EST) that cou ...

To attempt to gain an understanding of the molecular underpinnings of disease, many researchers have turned to expression profiling of genes during various stages of host recognition, entry, invasive growth, and host responses. While these studies have proven valuable, a deeper level of k ...

This chapter describes how a pipeline for the analysis of expressed sequence tag (EST) data can be �implemented, based on our previous experience generating ESTs from Trichoderma spp. We focus on key steps in the workflow, such as the processing of raw data from the sequencers, the clustering of ESTs, ...

Fusarium graminearum, the causal agent of head blight of wheat, was the third filamentous fungus to have a completed genome sequence. Since the release of the genome sequence in 2003, F. graminearum has become a model for studies of genomics and transcriptomics, mycotoxins, fungal population ...

Advances in genome sequencing technologies have facilitated production of a wealth of fungal data; within the last 5 years, experimental costs and labor have diminished, shifting the production bottleneck from genomic data generation to data analysis. Genome sequences and microar ...

Since the first fungal genome was sequenced in 1996, sequencing technologies have advanced dramatically. In recent years, it has become possible to cost-effectively generate vast amounts of DNA sequence data using a number of cell- and electrophoresis-free sequencing technologie ...

Advances in sequencing technologies have fundamentally changed the pace of genome sequencing projects and have contributed to the ever-increasing volume of genomic data. This has been paralleled by an increase in computational power and resources to process and translate raw seque ...

Our understanding of RNA has evolved over the last 20 years from the initial concept that RNA is simply an intermediate in protein synthesis or a structural component maintaining and expressing genetic information. Subsequently, the non-coding RNAs have attracted huge interest and have ...

The use of RNase P ribozyme (M1GS catalytic RNA) for inhibition of murine cytomegalovirus (MCMV) propagation in mice is described in this chapter. General information about RNase P based technology is included and followed by detailed protocols focused on (1) construction and in vitro clea ...

Triplex ribozymes allow for the individual activity of multiple trans-acting ribozymes producing higher target cleavage relative to tandem-expressed RZs. A triplex expression system based on a single hairpin ribozyme for the multiple expression (multiplex) vectors can be engi ...

Aptamers are ssDNA, RNA, or modified nucleic acids, usually consisting of short strands of oligonucleotides. Aptamers have the ability to bind specifically to a range of targets, from small organic molecules to proteins. However, by using cell-based aptamer selection, we have developed a s ...

Aptamers are high-affinity oligonucleotides which can be selected from large random libraries by systematic evolution of ligands by exponential enrichment (SELEX) protocols, with affinities and specificities comparable or better than antibodies. The SELEX protocols comp ...

Small interfering RNAs (siRNAs) have emerged as a powerful technique for sequence-specific gene silencing in a wide variety of organisms. However, it has been shown that in vitro transcribed siRNA can activate innate immunity through RIG-1 helicase, a cytosolic sensor of viral RNA. We recen ...

Since decades, the main goal of tumor immunologists has been to increase the capacity of the immune system to mediate tumor regression. Considerable progress has been made in enhancing the efficacy of therapeutic anticancer vaccines. First, dendritic cells (DCs) have been identified as t ...

Over the past decade, immunotherapy has emerged as a promising alternative form of cancer treatment with the potential to eradicate tumour metastasis. However, its curative potential is in general limited by peripheral tolerance mechanisms and the elimination of self-reactive T cel ...

Adoptive transfer of T cells with restricted tumor specificity provides a promising approach to immunotherapy of cancers. However, the isolation of autologous cytotoxic T cells that recognize tumor-associated antigens is time consuming and fails in many instances. Alternative ...

RNA modifications impact numerous cellular processes such as pre-mRNA splicing and protein synthesis. The elucidation of the mechanisms by which these modifications impact cellular processes necessitates the ability to both detect and quantify the presence of these modificat ...

In adults, the bone marrow compartment contains hematopoietic stem cells (HSCs) which can differentiate into progenitors with more restricted lineage potential and generate all cellular elements of the blood. HSCs for stem cell transplantation can be obtained by bone marrow collec ...