Molecular cloning of cDNA sequences transcribed from mouse liver endoplasmic reticulum poly(A)mRNA.
Poly(A)mRNA was isolated from the endoplasmic reticulum (ER) of male BALB/c mice and used as the template for cDNA synthesis. Double-stranded (ds) cDNA was cloned in a bacterial plasmid and 21 clones were selected for further study. Six clones have been identified by hybrid selection of mRNA, translation of the mRNA in a template-dependent system, polyacrylamide gel electrophoresis and specific immunoprecipitation. Four code for the major urinary protein of the mouse (MUP), one for serum albumin and one for alpha 1-antitrypsin. A seventh codes for an unidentified polypeptide with Mr = 27000. Initial-rate hybridisation kinetics of immobilised cloned sequences with end-labelled mRNA showed that serum albumin mRNA is most abundant in the population of mRNA species present in the endoplasmic reticulum fraction of female BALB/c liver. MUP mRNA is also very abundant. The alpha 1-antitrypsin mRNA and the mRNA specifying the 27000-dalton protein are about 5 times less abundant than serum albumin mRNA on a molar basis.
D000818:Animals; D003001:Cloning, Molecular; D004247:DNA; D004262:DNA Restriction Enzymes; D004721:Endoplasmic Reticulum / Q000032:analysis*; D008099:Liver / Q000648:ultrastructure; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D011061:Poly A / Q000235:genetics*; D011506:Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D012709:Serum Albumin / Q000235:genetics; D000515:alpha 1-Antitrypsin / Q000235:genetics
D011506:Proteins; D012333:RNA, Messenger; D012709:Serum Albumin; D000515:alpha 1-Antitrypsin; D011061:Poly A; D004247:DNA; D004262:DNA Restriction Enzymes