The isolation of cDNA clones for human apolipoprotein E and the detection of apoE RNA in hepatic and extra-hepatic tissues.

摘要: We have isolated cDNA clones coding for apolipoprotein E (apoE) from a library prepared adult human liver mRNA. Mixtures of 128 different oligonucleotides, 17 residues long were synthesised to be complementary regions the mRNA corresponding amino acids 1-6 and 151-156. Five independent apoE selected by direct screening 5000 recombinants with two oligonucleotide mixtures. Two overlapping contain 3'-untranslated sequence, entire sequence an additional 30 bases 5' terminus mature protein. The DNA has been determined spanning known sites acid substitutions which account observed protein polymorphism apoE. Using as probes in Northern blot analysis total kidney RNAs leucocyte poly(A)+ RNA we detected single species 1.2 kb larger RNA. level expression is approximately 10% that while sequences leucocytes less than 1% liver.