[Transcription start site determination in Stylonychia lemnae tubulin genes].
Contemporary experimental literature concerning transcription regulation study contains a lot of inconsistencies between data on precise transcription start site position in tubulin genes of hypotrychious ciliates. We have revealed that the reason of these inconsistencies lies in the methods applied by scientists. In present study, the basic methods of transcription start site identification were analyzed. The modification of transcription start site identification method based on the analysis of sequencing and primer extension reactions products on the same PAA gel was elaborated using Stylonychia lemnae tubulin genes as a model object. The use ofnon-phosphorylated primers in sequencing reaction (in accordance with standard procedure) leads to the appearance of band shifting which causes mistakes in results evaluation. This band shifting can be eliminated by the use of primers phosphorylated by non-radioactive phosphorus. In present study we applied this method to redetermine transcription start sites in alpha1-, alpha2-, beta1- and beta2-tubulin genes of S. lemnae.
Pimenov
A Iu
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D000818:Animals; D001483:Base Sequence; D016798:Ciliophora / Q000235:genetics*; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D017125:Genes, Protozoan; D015800:Protozoan Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis; D016053:RNA, Protozoan / Q000096:biosynthesis; D017422:Sequence Analysis, DNA / Q000379:methods*; D024363:Transcription Initiation Site; D014404:Tubulin / Q000235:genetics*
D016054:DNA, Protozoan; D015800:Protozoan Proteins; D012333:RNA, Messenger; D016053:RNA, Protozoan; D014404:Tubulin