cry1Aa lacks stability elements at its 5'-UTR but integrity of its transcription terminator is critical to prevent decay of its transcript.
We analyzed the influence of the 5' and 3' untranslated regions of the Bacillus thuringiensis cry1Aa on its mRNA stability. Although the cry1Aa gene has a stable transcript (8 min), its 5' UTR did not provide stability to the reporter gene uidA. Stability of cry1Aa could be increased to 40 min by addition of an SP82 stability element at the 5' UTR, suggesting that once the 5' and 3' ends were protected initiation of decay could be effectively blocked. We generated mutations in the transcription terminator and found that changes that reduced the stability of the stem, a larger loop, or elimination of the U-trail sharply decreased the half-life of the transcript. Therefore, unlike some stable bacterial transcripts, cry1Aa lacks special features at the end 5' to prevent decay, but its terminator is the main determinant of its stability.
Ramírez-Prado
Jorge Humberto
JH
Departamento de Ingeniería Genética, Centro de Investigación y de Estudios Avanzados, Unidad Irapuato, México.
Martínez-Márquez
Eva Isabel
EI
Olmedo-Alvarez
Gabriela
G
D020121:5' Untranslated Regions / Q000737:chemistry / Q000235:genetics; D001413:Bacillus thuringiensis / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001427:Bacterial Toxins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D015152:Blotting, Northern; D018388:Codon, Terminator / Q000235:genetics; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D008957:Models, Genetic; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000235:genetics; D012323:RNA Processing, Post-Transcriptional; D020871:RNA Stability; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014158:Transcription, Genetic / Q000235:genetics
D020121:5' Untranslated Regions; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D018388:Codon, Terminator; D012329:RNA, Bacterial; D011993:Recombinant Fusion Proteins; D005966:Glucuronidase