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Revista Latinoamericana de Microbiología

2000, Number 3

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Microbiología 2000; 42 (3)

Effect of λ tI point mutations on T3 RNA polymerase-mediated transcription termination

Cisneros B, Bermúdez CRM, Sánchez A, Montañez C

Language: Spanish
References: 32
Page: 95-100
PDF size: 87.62 Kb.


ABSTRACT

The terminator tI, located approximately 280 nucleotides beyond the int gene of bacteriophage λ, is considered an intrinsic terminator due to its GC-rich region of dyad symmetry followed by a T stretch. In order to study the effect of tI structure in T3 RNA polymerase-mediated transcription termination, three different point mutations tI1, tI2 and tI3 were analyzed. DNA fragments containing the tI mutations were cloned into the pBS+ vector for in vitro transcription studies. All the tI mutations mapping in the tI stem-loop structure decrease the in vitro transcription termination of tI from 85% for the wild type terminator to 46-60% using the T3 RNA polymerase.


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