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- W1625101491 abstract "This chapter presents extension inhibition to the study of translation initiation complexes. The chapter also describes the method of using bacteriophage T4 gene 32 mRNA. Early experiments showed that ribosomes, in the presence of fMet-tRNAMetf could properly select translation initiation regions of bacteriophage RNA and protect these regions from ribonuclease digestion. This chapter subsequently demonstrated that a 70S–fMet-tRNAMetf complex, bound to the coat protein initiation site on Q β plus strand RNA, cannot be dislodged by Q β replicase engaged in negative strand synthesis. It was reasoned that specific binding of ribosomes (30S or 70S) to mRNA might lead to pausing or termination of reverse transcriptase when a bound ribosome is encountered. In extension inhibition, a 5'-32P-end-labeled oligo deoxyribonucleotide, complementary to a region on the mRNA that is 3' to the initiation codon, is annealed to either total cellular RNA or to a purified transcript. This [32P]oligonucleotide-mRNA hybrid is then incubated with ribosomes, and the complexes analyzed by primer extension." @default.
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- W1625101491 date "1988-01-01" @default.
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- W1625101491 title "[27] Extension inhibition analysis of translation initiation complexes" @default.
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- W1625101491 doi "https://doi.org/10.1016/s0076-6879(88)64058-4" @default.
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