We designed a new inverse PCR protocol combined with switching mechanism at 5' end of RNA transcript (SMART) technology, and applied it to the cloning of teleost corticotropin-releasing hormone precursor cDNA. Due to the advantages of both techniques, this method can efficiently amplify the complete 5'- and 3'-ends of cDNA in a single reaction, and might prove to be an alternative to the conven...

In 1983, we described a simple protocol to synthesize cDNA by a combination of reverse transcriptase, RNAseH and DNA polymerase I to generate large plasraid libraries (1). This protocol has now been modified to a simple one tube reaction with bluntended cDNA as the final product that can be cloned into bacteriophage lambda or plasmids. Preparation of the cDNA takes about 4 hours; a library in l...

An efficient cDNA amplification procedure is described for determining of the 5' and 3' ends of mRNAs and cloning full-length cDNAs. In this approach, a double-stranded (ds) adaptor is ligated to both ends of a library of ds cDNA by T4 DNA ligase. This adaptor-ligated ds cDNA is then used to selectively amplify 5'- or 3'-cDNA fragments by PCR with a combination of gene-specific and adaptor-spec...

Background: Recent researches have demonstrated that human T-cell immunoglobulin mucin 1 (TIM-1) glycoprotein plays important roles in regulation of autoimmune and allergic diseases, as well as in tumor immunity and response to viral infections. Therefore,targeting TIM-1 could be a potential therapeutic approach against such diseases. Objectives: In this study, we aimed to express TIM-1 protein...

1 Identification of di (2-ethylhexyl) phthalate-induced carboxylesterase 1 in C57BL/6 mouse liver microsomes: Purification, cDNA cloning, and baculovirus-mediated expression.

background: recent researches have demonstrated that human t-cell immunoglobulin mucin 1 (tim-1) glycoprotein plays important roles in regulation of autoimmune and allergic diseases, as well as in tumor immunity and response to viral infections. therefore,targeting tim-1 could be a potential therapeutic approach against such diseases.objectives: in this study, we aimed to express tim-1 protein ...

The complete amino acid sequence of the receptor for organic calcium channel blockers (CaCB) from rabbit lung has been deduced by cloning and sequence analysis of the cDNA. Synthetic RNA derived from this cDNA induces the formation of a functional CaCB-sensitive high voltage activated calcium channel in Xenopus oocytes.

1. The Importance of the Mouse Genome and Transcriptome Study 2. The History of Mouse Genetics 2.1. The Beginning of Mouse Genetics and the Establishment of Inbred Strains 3. The Determination of Mouse Genome Sequences 4. A Full-Length Mouse cDNA Project 4.1. From the EST Project to the Full-Length cDNA Project 4.2. Findings from Genome and cDNA Sequences 4.3. Functional Annotation of Mouse cDN...

One of the most basic techniques in biomedical research is cDNA cloning for expression studies in mammalian cells. Vaccinia topoisomerase I-mediated cloning (TOPO cloning by Invitrogen) allows fast and efficient recombination of PCR-amplified DNAs. Among TOPO vectors, a pcDNA3.1 directional cloning vector is particularly convenient, since it can be used for expression analysis immediately after...