Construction and partial characterization of a human liver cDNA library.
Total messenger RNA (mRNA) was isolated from adult human liver and copied to give complementary DNA (cDNA) with reverse transcriptase. Double stranded cDNA was cloned by GC tailing into the PstI site of the plasmid pAT153; approximately 2000 recombinant clones were isolated. The sensitivity of the bacterial cells to ampicillin was used as a marker to study the stability of the library during consecutive overnight growth in mixed liquid culture. A significant change in the composition of the library was observed after three overnight cultures. A heterologous rat albumin cDNA recombinant and two specific oligonucleotides were used to screen the library, and clones containing sequences of the human mitochondrial rRNA- and the alpha-1 antitrypsin-genes were isolated.
D000418:Albumins / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D006801:Humans; D008099:Liver / Q000378:metabolism*; D008930:Mitochondria, Liver / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics; D051381:Rats; D000515:alpha 1-Antitrypsin / Q000235:genetics
D000418:Albumins; D012333:RNA, Messenger; D012335:RNA, Ribosomal; D000515:alpha 1-Antitrypsin; D004247:DNA