An expression system for human apolipoprotein B100 in a rat hepatoma cell line.
We have designed an in vitro expression system for human apolipoprotein (apo) B. A full-length human apoB minigene was constructed from cDNA and genomic apoB clones and inserted into a vector where its expression was directed by the cytomegalovirus promoter. The apoB minigene was expressed in a rat hepatoma cell line, McA-RH7777. Human apoB100, which is the ligand for the low density lipoprotein receptor, was secreted in low density lipoprotein or very low density lipoprotein particles, depending on the composition of the medium. A protein with the mobility of apoB48, a structurally related protein involved in cholesterol metabolism, was also produced from the human apoB minigene. This in vitro expression system for human apoB will enable investigators to identify which domains of this protein are involved in processes such as lipoprotein assembly and secretion. This system should also allow investigators to identify definitively the domain in apoB that enables the protein to bind to the low density lipoprotein receptor.
Blackhart
B D
BD
Gladstone Foundation Laboratories for Cardiovascular Disease, University of California, San Francisco 94140-0608.
Yao
Z M
ZM
McCarthy
B J
BJ
D000818:Animals; D053299:Apolipoprotein B-100; D001055:Apolipoproteins B / Q000096:biosynthesis / Q000235:genetics*; D002460:Cell Line; D015870:Gene Expression; D005796:Genes; D005822:Genetic Vectors; D006801:Humans; D008114:Liver Neoplasms, Experimental / Q000378:metabolism; D051381:Rats; D011973:Receptors, LDL / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000378:metabolism*
D053299:Apolipoprotein B-100; D001055:Apolipoproteins B; D011973:Receptors, LDL; D011993:Recombinant Fusion Proteins