Messenger RNA for hydroxyindole O-methyltransferase (EC 2.1.1.4) was partially purified from poly(A)+ RNA isolated from bovine pineal glands by sucrose density gradient centrifugation. The enriched mRNA was used to prepare a cDNA library by use of expression vector lambda gt11. The library was screened with monoclonal antibodies to the enzyme, and three cDNA clones were isolated. These cloned c...

Vaccines that induce mosquito-killing (mosquitocidal) activity could substantially reduce the transmission of certain mosquito-borne diseases, especially vaccines against African malaria vectors, such as the mosquito Anopheles gambiae. To generate and characterize antimosquito immunity we immunized groups of mice with two individual A. gambiae midgut cDNAs, Ag-Aper1 (a secreted peritrophic matr...

We have developed a PCR-based method for rapid and effective screening of arrayed cDNA libraries. This strategy directly addresses the limitations of conventional hybridization-based schemes and provides a more rapid, cost-effective, and sensitive method compatible with large-scale and routine cDNA clone recovery. To prepare arrayed libraries, 1-2 x 10(6) cDNA clones were propagated as individu...

target sequence is correct, the target sequence is amplified, and if the primer pair is in the wrong orientation relative to the amplified cDNA inserts, the target sequence is not amplified. This simple, linear pre-amplification of cDNAs in the library has the potential for wide use. This approach requires not only far fewer starting materials than conventional PCR cloning but also circumvents ...

Fructose-1,6-bisphosphatase (EC 3.1.3.11) catalyzes the dephosphoric reaction of Fru-1,6-bis P to Fru-6-P and Pi. Two isozymes exist in higher plants, one in the cytosol and the other in the chloroplast. The latter is a pivotal enzyme for photosynthetic carbon dioxide assimilation. It is active in the light but almost inactive in darkness. The lightmediated activation of the enzyme is attribute...

We here describe a convenient system for the production of recombinant adenovirus vectors and its use for the construction of a representative adenovirus-based cDNA expression library. The system is based on direct site-specific insertion of transgene cassettes into a replicating donor virus. The transgene is inserted into a donor plasmid containing the viral 5' inverted terminal repeat, the co...

Mammalian cytochrome c oxidase (COX) is composed of three catalytic, on mitochondrial DNA encoded subunits, and ten nuclear encoded subunits, which have been ascribed a regulatory function (1). At least three nuclear encoded subunits occur in tissue-specific isoforms (Via, Vila and VIE). The liverand hearttype cDNA has been described for rat subunit Via (2) and bovine subunit Vm (3). The liver-...

In this study we successfully constructed a full-length cDNA library from Siberian tiger, Panthera tigris altaica, the most well-known wild Animal. Total RNA was extracted from cultured Siberian tiger fibroblasts in vitro. The titers of primary and amplified libraries were 1.30×10(6) pfu/ml and 1.62×10(9) pfu/ml respectively. The proportion of recombinants from unamplified library was 90.5% and...

The Prostate Expression Database (PEDB) is a curated relational database and suite of analysis tools designed for the study of prostate gene expression in normal and disease states. Expressed Sequence Tags (ESTs) and full-length cDNA sequences derived from more than 40 human prostate cDNA libraries are maintained and represent a wide spectrum of normal and pathological conditions. Detailed libr...

Collection of full-length genes requires libraries with full-length cDNA insert, large-scale sequencing, library assessment, and high-speed sequence clustering. Here we focus on computational methods, such as newly developed computer programs, since our experimental methods had been published previously. Our purpose is the collection of full-length cDNAs, therefore the proportion of full-length...