Novel membrane-bound reporter molecule for sorting high producer cells by flow cytometry.

We developed a membrane bound reporter and selection molecule for sorting by fluorescence activated cell sorting (FACS) of cells producing a protein of interest. This molecule is composed of a transmembrane (TM) domain, fused on its extracellular end to a biotin mimetic peptide (BMP) and on its intracellular side to puromycin N-acetyl transferase (PAC). In this format BMP is displayed on the cell membrane surface and PAC faces the cell cytoplasm.

The low-density lipoprotein receptor plays a role in the infection of primary human hepatocytes by hepatitis C virus.

Background/aims: The direct implication of low-density lipoprotein receptor (LDLR) in hepatitis C virus (HCV) infection of human hepatocyte has not been demonstrated. Normal primary human hepatocytes infected by serum HCV were used to document this point.

Methods: Expression and activity of LDLR were assessed by RT-PCR and LDL entry, in the absence or presence of squalestatin or 25-hydroxycholesterol that up- or down-regulates LDLR expression, respectively.

A novel lipid-conjugated marker for the direct labeling of the schistosomulum membrane.

Introduction of the fluorescent group lissamine rhodamine B into the surface of schistosomula of S. mansoni was achieved by brief incubation of the worms with liposomes carrying the lipid bound fluorophore in their bilayers. The liposomes were made of egg lecithin (PC) and lissamine-phosphatidylethanolamine (lissamine-PE).

Isolation and partial characterization of the tegumental outer membrane of schistosomula of Schistosoma mansoni.

Separation of the external membranes from freshly converted mechanical schistosomula of Schistosoma mansoni was achieved by osmotic shock under hypertonic conditions, followed by mechanical shearing and ultracentrifugation. Prior to treatment, the schistosomula were surface labeled by introduction of N-DNP-epsilon-aminocaproylphosphatidylethanolamine molecules into their lipid bilayer followed by anti-DNP antibodies and stained with either 125I-protein-A or ferritin labeled secondary anti-DNP antibodies. This label provided a membrane marker by which the purity of the preparation could be assessed at each stage.

Acetylcholinesterase of Schistosoma mansoni: antigenic cross-reactivity with Electrophorus electricus and its functional implications.

Acetylcholinesterase (AChE) of the parasite Schistosoma mansoni cross-reacts with rabbit antibodies against AChE from the electric eel. Cross-reactivity was demonstrated by radioimmunoassay with sonicate, as well as soluble, preparations from various stages of the parasite life cycle. On sucrose density gradient, the parasite enzyme migrates as an 7.