In vivo transcription initiation and termination sites of an alpha-amylase gene from Bacillus amyloliquefaciens cloned in Bacillus subtilis.
The alpha-amylase gene, originally isolated by molecular cloning from chromosomal DNA of Bacillus amyloliquefaciens, is efficiently expressed from its own promoter in a Bacillus subtilis host when present in the multicopy plasmid vector pUB110. The flanking regions of this gene were sequenced and the ends of the in vivo-generated messenger RNA were mapped by the S1 procedure. Outside the coding sequence, the mRNA for alpha-amylase contains about 30 nucleotides at the 5' end and 51 nucleotides at the 3' end. The promoter region has -10 sequence TAAAAT starting eleven nucleotides upstream from the transcription start point, pppU, and the -35 hexanucleotide TTGTTA is separated from it by 16 nucleotides. As indicated by its sequence, the terminator is bidirectional and of the rho-independent kind, and the mRNA can form a long hairpin structure at the very 3' end. The 3' terminus of the transcript does not seem to include a U stretch, although the DNA template codes for U3AU6 at the 3' end of the hairpin sequence. The bulk of the amylase mRNA does not contain any 3'-terminal poly(A).
D001407:Bacillus / Q000235:genetics*; D001412:Bacillus subtilis / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D009876:Operon; D011061:Poly A / Q000235:genetics; D012329:RNA, Bacterial / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic; D000516:alpha-Amylases / Q000235:genetics*
D012329:RNA, Bacterial; D012333:RNA, Messenger; D011061:Poly A; D000516:alpha-Amylases