Polyadenylylation in mycobacteria: evidence for oligo(dT)-primed cDNA synthesis.
The ability of mRNA to direct synthesis of cDNA in the presence of oligo(dT) was analysed using a novel application of fluorescein-11-dUTP incorporation into cDNA by reverse transcriptase. Evidence is provided for the first time that a majority of the mycobacterial mRNA pool is polyadenylylated. mRNA transcripts of hsp65 were also amplified with specific primers from the oligo(dT)-primed cDNA preparation in Mycobacterium bovis BCG, M. smegmatis and M. vaccae. Furthermore, PCR amplication of cDNAs for genes entD, entC and trpE2 from M. bovis BCG yielded the expected products when reverse transcription was primed with oligo(dT), suggesting that polyadenylylation is a general phenomenon in mycobacteria.
Adilakshmi
Tadepalli
T
Department of Biological Sciences, University of Hull, Hull HU6 7RX, UK1.
Ayling
Peter D
PD
Department of Biological Sciences, University of Hull, Hull HU6 7RX, UK1.
Ratledge
Colin
C
Department of Biological Sciences, University of Hull, Hull HU6 7RX, UK1.
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015139:Blotting, Southern; D018834:Chaperonin 60; D018833:Chaperonins / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary / Q000096:biosynthesis*; D003856:Deoxyuracil Nucleotides / Q000378:metabolism; D005452:Fluoresceins / Q000378:metabolism; D009161:Mycobacterium / Q000235:genetics* / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000378:metabolism*; D011061:Poly A / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012329:RNA, Bacterial / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D018834:Chaperonin 60; D018076:DNA, Complementary; D003856:Deoxyuracil Nucleotides; D005452:Fluoresceins; D009838:Oligodeoxyribonucleotides; D012329:RNA, Bacterial; D012333:RNA, Messenger; C074119:heat-shock protein 65, Mycobacterium; C027903:oligo (dT); C069632:fluorescein-11-deoxyuridine triphosphate; D011061:Poly A; D018833:Chaperonins