A cDNA library was constructed from firefly (Photinus pyralis) lantern poly(A)+ RNA, using the Escherichia coli expression vector lambda gt11. The library was screened with anti-P. pyralis luciferase (Photinus luciferin:oxygen 4-oxidoreductase, EC 1.13.12.7) antibody, and several cDNA clones expressing luciferase antigens were isolated. One clone, lambda Luc1, contained 1.5 kilobase pairs of cD...

A new approach for simultaneously amplifying the 5' and 3' ends of a desired cDNA is described. The method combines the template-switching effect with inverse PCR, which generates the flanking 5' and 3' regions of a certain cDNA with very low or no background. It requires only minimal amounts of total RNA for the synthesis of first-strand cDNA, while the same cDNA can be used to amplify flankin...

Cardiac pacemaking offers a unique opportunity for direct gene transfer into the heart. An experimental system was developed to assay the effects of transferring the human b 2 adrenergic receptor ( b 2 AR) under in vitro, ex vivo, and finally in vivo conditions. Constructs encoding either b 2 AR or LacZ were used in chronotropy studies with isolated myocytes, and transplanted as well as endogen...

We have developed a fast and general method to obtain an enriched, full-length cDNA expression library with subtractively enriched cDNA fragments. The procedure relies on RecA-mediated triple-helix formation of single-stranded cDNA fragments with a double-stranded cDNA plasmid library. The complexes were then captured from the solutions using the digoxigenin-antidigoxigenin paramagnetic beads f...

PURPOSE The purpose of our study was to determine whether the amounts of circulating DNA (cDNA) could discriminate between NSCLC patients and healthy individuals and assess its value as a prognostic marker of this disease. METHODS We conducted a study of 309 individuals and the cDNA levels were assessed through real-time PCR methodology. RESULTS We found increased cDNA levels in NSCLC patie...

The gene encoding the human basic fibroblast growth factor (hbFGF) has been already chemically-synthesized and cloned in pET-3a expression vector (Pasteur Institute of Iran). In the present study, we compared the level of expression of this synthetic hbFGF and its related cDNA in Escherichia coli. The pBR322-cDNA of hbFGF supplied by Dr. Seno (from Molecular Biology Dept, Okaido prefectural uni...

Several groups have isolated cDNA clones encoding different thyroid hormone binding proteins, although in each case only one type of cDNA was characterized for a given tissue (1-5). We have found three different cDNAs in libraries prepared from mouse skeletal muscle tissue at different stages of development and from the mouse C2 muscle cell line. Two different probes were used to screen these l...

background: pentavalent antimonials are the first line drugs for the treatment of leishmaniasis. unresponsiveness of leishmania spp. to antimonial drugs is a serious problem in some endemic areas. investigations on molecular mechanisms involved in drug resistance are essential for monitoring and managing of the disease. cal-cineurin is an essential protein phosphatase for number of signal trans...