Recent studies have shown that the microRNA (miRNA) pathway is an evolutionarily conserved endogenous pathway that is important for normal development. Mature miRNAs are excised from precursors in a stepwise process and subsequently incorporated into an RNA-induced silencing c ...
MicroRNAs (miRNAs), small single-stranded regulatory RNAs capable of interfering with intracellular messenger RNAs (mRNAs) that contain either complete or partial complementarity, are useful for the design of new therapies against cancer polymorphism and viral mutation. Nu ...
Prediction of microRNA (miRNA) candidates using computer programming has identified hundreds and hundreds of genomic hairpin sequences, of which, the functions remain to be determined. Because direct transfection of hairpin-like miRNA precursors (pre)-miRNAs in mammalian c ...
As more and more genes are cloned and sequenced, it is apparent that nearly all genes are related to other genes. Similar genes are grouped into families, such as the collagen and globin gene families. There are also gene superfamilies. Gene superfamilies are composed of genes that have areas of high ho ...
The Lambda ZAP� vectors have been designed to allow in vivo excision and recircularization of the cloned insert and phagemid sequences contained within the λ vector (1). Once a Lambda ZAP library is constructed and amplified, putative clones or the λ library itself may be excised into the phagemid f ...
The starting material for any representative plant cDNA library is a supply of good-quality messenger RNA from the plant tissue of choice. Extraction of RNA can be made from several grams of tissue or as little as 50 mg. However, large samples are generally more representative of the genes expressed in a ...
It is estimated that 50,000–100,000 genes are expressed across the cell types of higher eukaryotes. As a consequence of differential expression (either regionally, temporally, or environmentally specific), a large proportion of all transcripts (approx 40–45%) represent low-abun ...
The way cDNA libraries are used for finding genes of interest depends on the overall strategy adopted. Therefore, before exploring the options for cDNA library access, we briefly delineate these strategies and the place of cDNA libraries within them.
The accumulation and analysis of expressed sequence tags (ESTs) are now an important component of genome research (1). An EST is a fragment of a sequence from a cDNA clone that corresponds to an mRNA. ESTs have applications in the discovery of new human genes, mapping of the human genome, and identificat ...
The Human cDNA Database (HCD) is a repository for human cDNA sequences and related data that is curated and maintained at The Institute for Genomic Research (TIGR). The foundation of the database is ∼160,000 partial cDNA sequences that have been generated by TIGR and Human Genome Sciences (HGS). Th ...
Since the first report on cDNA cloning in 1972 (1), this technology has been developed into a powerful and universal tool in the isolation, characterization, and analysis of both eukaryotrc and prokaryotic genes. But the conventional methods of cDNA cloning require much effort to generate a li ...
In the study of biological systems, often the amount of starting material available for molecular analysis is limiting. In some situations, only a few cells in a tissue are expressing genes of interest or the tissue is in limited supply. A cDNA library from the targeted cells is preferable to a library co ...
Analysis of the data generated from cDNA sequences, an important step in the final stages of any sequencing project, can provide insights into gene structure and function as well as help to direct the experimental approaches to obtaining the sequence of a full-length clone even when only prelim ...
Cellular phenotypes resulting from decreased function of a specific gene are manifested as recessive, since they are suppressed in the presence of a normal allele of the corresponding gene. The powerful gene-transfer techniques, which have played a key role in the studies of dominant pheno ...
The phage λgt1.1 system has become increasingly popular for expression of cDNAs or genomic DNAs either in phage plaques or in bacteria lysogenized with recombinant phages (1,2). It offers the advantages of high cloning efficiency, high-level expression, the relative stability of β-gala ...
One of the most powerful ways to gain novel insights into a particular protein’s cellular functions is to identify other proteins with which it specifically interacts. Traditionally, this problem has been approached by use of protein biochemistry, involving purification of interact ...
MicroRNAs (miRNAs) form a large class of small regulatory RNAs in eukaryotes. Although they share a common processing pathway and certain structural features, in general, there is no detectable sequence similarity among miRNAs from a given organism. On the other hand, many miRNAs are members ...
MicroRNA biogenesis occurs in several steps from their precursors having irregular hairpin structures. The highly variable architecture of these stem-and-loop structures, which have terminal loops of various sizes and diverse structure destabilizing motifs present in their ...
The discoveries of microRNAs (miRNAs), a class of noncoding RNAs that can regulate gene expression by translational repression, have opened a new avenue on gene modulation in mammalian cells. Conceivably, this highly effective method of modulation of gene expression would be exploited ...
The recently discovered microRNAs (miRNAs) are a large family of small regulatory RNAs that have been implicated in controlling diverse pathways in a variety of organisms (1,2). For posttranscriptional gene silencing, one strand of the miRNA is used to guide components of the RNA interfere ...
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