Simian virus 40 as a vector: recombinant viruses expressing individual polyoma T antigens.

We constructed simian virus 40 (SV40)/polyomavirus recombinants by replacing in SV40 the T antigen coding region with polyoma early region sequences, either cDNAs encoding small, middle or large T antigen or the wild-type sequence coding all three proteins. The recombinants maintained the SV40 late region and origin of replication and were propagated in COS cells yielding recombinant virus preparations with titers of 10(6)-10(7) infectious particles per milliliter. These viruses were characterized in productive infections of COS cells by analyzing early and late mRNA levels and by following synthesis of polyoma early proteins.

Rapid detection of the main human plasma glycoproteins by two-dimensional polyacrylamide gel electrophoresis lectin affinoblotting.

Glycoprotein modifications in the glycan moiety can occur in diseases such as cancers, inflammatory processes and alcoholism. We combined high-resolution two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) with lectin affinoblotting in order to establish the normal human plasma glycoprotein map. Human plasma proteins were separated by mini 2-D PAGE (7 x 9 cm), transferred onto polyvinylidene difluoride membranes and incubated with biotinylated lectins.

Alcohol flushing, patch test, and ADH and ALDH genotypes in Brazilian ethnic groups.

Self reports of flushing reaction after drinking, cutaneous sensitivity to alcohol (patch test), and genotypic determination of ADH2, ADH3, and ALDH2 were studied in 53 Brazilian volunteers of different ethnic groups. Genotypes were determined using single-strand conformation polymorphism in discontinuous buffer electrophoresis. Analysis of the results indicated several cases of a reported flushing reaction among ALDH2 1/1 individuals, while all but 2 cases of ALDH2 heterozygotes reported a flushing reaction.

Analysis of glycoproteins separated by two-dimensional gel electrophoresis using lectin blotting revealed by chemiluminescence.

The carbohydrate structures of blotted glycoproteins can be analyzed by probing with lectins. The objective of the present work was to optimize the lectin blotting of human plasma glycoproteins separated by two-dimensional gel electrophoresis and the detection by the sensitive chemiluminescence method. The proposed detection method was found to be ten times more sensitive than a standard colorimetric reaction.

LAP (NF-IL-6), a tissue-specific transcriptional activator, is an inhibitor of hepatoma cell proliferation.

During postnatal liver development, LAP (NF-IL-6, C/EBP beta) expression and hepatocyte proliferation are mutually exclusive. In addition to transactivating liver-specific genes, LAP, but not C/EBP alpha, arrests the cell cycle before the G1/S boundary in hepatoma cells. LIP, a liver-inhibitory protein, which is translated from LAP mRNA lacking the activation domain of LAP, is not only ineffective in blocking hepatoma cell proliferation but also antagonizes the effect of LAP on the cell cycle.