Cloning of alpha 2u globulin cDNA using a high efficiency technique for the cloning of trace messenger RNAs.
An extremely high-efficiency technique is described for cloning double-stranded (ds) cDNAs in Escherichia coli. The method, which uses two synthetic oligonucleotide linkers rather than one, results in approx. 200--500 recombinant clones per ng of ds cDNA. This technique was used to clone a cDNA comprising 95% of the full length of the mRNA of alpha 2u globulin, a male rat liver protein, which represents approx. 1% of hepatic messenger RNA. The cloned probe was applied to study the complex hormone controls of alpha 2u globulin mRNA in male and female rats.
D000510:Alpha-Globulins / Q000235:genetics*; D000818:Animals; D002369:Castration; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000235:genetics; D004262:DNA Restriction Enzymes / Q000378:metabolism; D003907:Dexamethasone / Q000494:pharmacology; D013196:Dihydrotestosterone / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D008297:Male; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats
D000510:Alpha-Globulins; D012333:RNA, Messenger; C013803:alpha 2u globulin; D013196:Dihydrotestosterone; D004958:Estradiol; D003907:Dexamethasone; D004247:DNA; D004262:DNA Restriction Enzymes