遗传学实验技术

Interactions between RNA-binding proteins and cis-acting elements in the 5′- and 3′-untranslated regions (UTRs) of transcripts are responsible for regulating essential biological activities, such as mRNA localization, mRNA turnover, and translation efficiency. This chapt ...

This chapter provides a manual for a laboratory-based short course to introduce the common techniques of solid-phase peptide synthesis (SPPS). The course provides students the opportunity to design and manually synthesize analogs of glutathione using relatively simple equipme ...

The focus of this chapter is the isolation and analysis of subsets of proteins or subproteomes. These methodologies can be applied to address fundamental questions in many areas of biological research. Protocols are provided for common techniques that include protein isolation from ma ...

Bioconjugates play an important role in several fields of biomolecular and biomedicinal sciences. Protein/polypeptide-based conjugates with covalently attached epitope peptides are considered as potential synthetic vaccine candidates and/or target antigens in affi ...

This chapter describes backbone amide linker (BAL) strategies for the Nα-Fmoc solid-phase synthesis of C-terminal modified peptides. Most solid-phase protocols for the assembly of such peptides have limited generality, because they rely on the Cα-carboxyl for attachment to the solid ...

The methodologies of high-throughput peptide synthesis are overviewed and discussed. Particular focus is given to the techniques applicable to laboratories with a limited budget. Automated solutions for synthetic problems are also discussed.

Cyclic peptides have been reported to bind to multiple, unrelated classes of receptor with high affinity. Owing to the robustness of amide bond chemistry, the ability to explore extensive chemical diversity by incorporation of unnatural and natural amino acids, and the ability to explore c ...

The use of specific quenched fluorescent substrates (QFS) provides a rapid and sensitive method to measure peptidase activity, and is readily adaptable to highthroughput screening of potential peptidase inhibitors. In this chapter, we discuss general considerations for the deve ...

Peptide nucleic acid (PNA) is a promising tool in biomedical research. PNA oligomers can be synthesized according to peptide synthesis protocols, but they hybridize to complementary RNA and DNA strands like oligonucleotides. The main hindrance to the use of PNA oligomers has been their poor ...

Over the past decade, a significant methodological development in peptide ligation strategies has been elaborated that now permits the assembly of peptides and proteins. Native chemical ligation (NCL) has been introduced to join synthetic unprotected peptides by using the chemos ...

Phosphotyrosyl (pTyr)-containing sequences in proteins serve important roles in cellular signal transduction. Often, synthetic pTyr-containing peptides based on cognate sequences surrounding these pTyr residues can exhibit pharmacologically useful properties of ...

Cell-penetrating peptides (CPPs) have served as vehicles for the delivery of different molecules and particles into cells. The efficiency and noninvasive nature of peptide-mediated cellular transduction provides a promising tool for biomedical research. Studies of cell-pen ...

Chimeric peptides are unnatural constructs consisting of bioactive compounds from at least two different peptide(s) and/or protein(s) or two sequences from differentparts of the same protein. Such multifunctional peptide combinations are prepared to enhance the biological ac ...

The scientific literature is full of new small molecules and larger peptides identified as potential pharmaceutical agents for a variety of diseases. The majority of these compounds, however, will never progress into the clinic because of poor oral absorption and low metabolic stabili ...

The modular nature of peptides can be exploited in the synthesis of chimeric sequences that combine diverse motifs in a single molecule. A theoretical consideration of the classification of peptides further expounds the multigeneric nature of peptide chimeras. Strategies for chime ...

The purpose of this chapter is to delineate strategic considerations and provide practical procedures to enable non-experts to synthesize peptides with a reasonable chance of success. This chapter focuses on Fmoc chemistry, which is now the most commonly employed strategy for solid ph ...

Amplified Differential Gene Expression (ADGE) and DNA microarray provides a new concept that the ratios of differentially expressed genes are magnified prior to detecting them. The ratio magnification is achieved with the integration of DNA reassociation and polymerase chain rea ...

An increasing number of biological and medical research questions depend on obtaining global views of gene expression. In this chapter, we will describe how oligonucleotide microarrays have been used to accomplish this goal. In particular, we will focus on the use of GeneChip arrays�, which p ...

Scientists routinely talk and write about gene expression and the abundance of transcripts, but in reality they extrapolate this information from the various measurements that a variety of different technologies provide. Indeed, there are many reasons why applying different tech ...

The Invader� assay is a homogeneous, isothermal, signal amplification system for the quantitative detection of nucleic acids. The assay can directly detect either DNA or RNA without target amplification or reverse transcription. It is based on the ability of Cleavase� enzymes to recogni ...