Epitope tagging is a method of expressing proteins whereby an epitope for a specific monoclonal antibody is fused to a target protein using recombinant DNA techniques. The aim of this study was to sub-clone the peroxisomal protein (PEP) cDNA into a mammalian expression vector leading to the formation of a  chimeric PEP-cDNA containing the FLAG epitope. The FLAG-PEP recombinant cDNA was construc...

We present a simple molecular indexing method for quantitative targeted RNA sequencing, in which mRNAs of interest are selectively captured from complex cDNA libraries and sequenced to determine their absolute concentrations. cDNA fragments are individually labeled so that each molecule can be tracked from the original sample through the library preparation and sequencing process. Multiple copi...

Chitinase cDNAs from Leucaena leucocephala seed]ings were cloned by PCR amplification with degenerate primers based on conseryed class I chitinase sequences and cDNA library screening. Two closely related chitinase cDNAs were sequenced and inferred to encode precursor proteins of 323 (KBI) and 326 (KB2) amino acids. Expression of the KB2 chitinase from a pET32a plasmid in Origami (DE3) Escheric...

The genes of top athletes are a valuable genetic resource for the human race, and could be exploited to identify novel genes related to sports ability, as well as other functions. We analyzed the expressed sequence tags from top half-pipe snowboarding athletes using the SMART complementary DNA (cDNA) library construction method to elucidate the characteristics of the athlete genome and the diff...

We present evidence of cross-hybridization artifact intrinsic to spotted single-dye cDNA microarrays as a result of cDNA containing 5’-end sequences of consecutive thymidine (dT) residues. These poly(dT) tracts result from the synthesis, via oligo (dT) primed reverse transcription, of expressed sequence tags (EST) cDNA from a polyadenylated mRNA template. Analysis of gene expression data from t...