Expression of normal and mutagenized apolipoprotein CII in procaryotic cells. Structure-function relationship.
A full-length human apo CII cDNA clone has been constructed by completing the 5' end of an incomplete cDNA with a 44 bp long synthetic oligonucleotide. This apo CII cDNA insert was cloned into the pSP19 expression vector and transcribed and translated in vitro. Its N-terminal signal sequence (23 amino-acid residues) was accurately cleaved during cotranslational translocation through endoplasmic reticulum membranes to yield the mature apo CII. Mature apo CII was expressed on a preparative scale as fusion protein apo CII-beta-galactosidase with the full-length apo CII cDNA integrated into the pUR291 vector. Furthermore it was expressed in E. coli transformed with the pKK233-2 apo CII clone. The preform was accurately processed by the host cell. C-Terminal apo CII deletion mutants generated by partial Bal31 digestion of the pKK233-2 apo CII vector yielded well-defined truncated apo CII polypeptides on a preparative scale which allowed the determination of the polypeptide domain responsible for the activation of the serum lipoprotein lipase.
Holtfreter
C
C
Institut für Physiologische Chemie, Universität zu Köln.
Stoffel
W
W
D053304:Apolipoprotein C-II; D001056:Apolipoproteins C / Q000096:biosynthesis / Q000097:blood / Q000235:genetics*; D015139:Blotting, Southern; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004274:DNA, Recombinant / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005796:Genes; D006801:Humans; D009154:Mutation; D009693:Nucleic Acid Hybridization; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D014158:Transcription, Genetic
D053304:Apolipoprotein C-II; D001056:Apolipoproteins C; D004274:DNA, Recombinant; D011994:Recombinant Proteins; D004247:DNA