Molecular Cloning of the Left-Terminal SmaI-D Fragment of Adenovirus Type 7 DNA

The left-terminal SmaI-D fragment (map position 0-13.5%) of adenovirus type 7 DNA has been cloned in the plasmid vector pBR322 by the homopolymeric tailings of poly (dG) and poly (dC). Two isolated clones were characterized by mapping the cleavage sites of restriction endonucleases and by sequencing the junction between viral insert and pBR322

RT-PCR法によるヒトサイトメガロウイルス前初期遺伝子mRNAの検出

We have established a method for detection of human cytomegalovirus (HCMV) immediate early gene (IE)-specific mRNA by reverse transcription polymerase chain reaction (RT-PCR). Using the IE-1 specific primer fragments designed for RT-PCR, we have amplified a 232 base-pair fragment which represents IE-1 mRNA transcribed from the IE-1 gene of HCMV AD169 strain. No DNA cross reactivities to other human herpes virus DNAs or human embryonic lung cell (MRC-5 line) DNA were observed. The RT-PCR assay indicated HCMV IE-1-specific mRNA in HCMV-infected MRC-5 cells and in peripheral blood specimens from one of 3 patients examined. The results indicate that RT-PCR will be readily applicable to rapid detection of HCMV mRNA and may be useful for diagnosis of active primary or recurrent HCMV infections