生物化学原理教学（杨荣武）Chapter41 Regrogrammed genetic decoding and translational pause.pptVIP

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生物化学原理教学（杨荣武）Chapter41 Regrogrammed genetic decoding and translational pause.ppt

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Chapter41 Regrogrammed genetic decoding translational pause Outline Regrogrammed genetic decoding Translational frameshifting Read-through Jump-translation translational intron The Incorporation of Sec and Pyl Translational pause (self-study) Read-through In Mo-MuLV RNA, gag and pol are in the same reading frame but separated by a termination codon. Translation of the polycistron is accomplished by suppression by a Gln- tRNA. Similarly, the RNA of tobacco mosaic virus contains a suppressible stop codon. The read through product contains the RNA polymerase domain. Sequence context determines whether suppression occurs at a given stop codon. A complex signal prevents normal stop codons from being suppressed. Both upstream and downstream of the suppressible stop codon are required for suppression.? Regions downstream seem more conserved than those upstream Termination suppression has the same effect as frameshifting: the production of two proteins, one longer than the other. The efficiency of termination suppression determines the stoichiometric ratio of the longer to the shorter product Jump translation translational intron Bacteriophage T4 gene 60 (18 k subunit of DNA Topoisomerase). By comparison between cDNA and AA sequences, 50nts of the coding region of the mRNA are skipped as the mRNA slips between two identical Gly codons (GGA) 50 nts apart, with close to 100% efficiency of bypass. A Gene 60- lacZ fusion is skipped with almost 100% efficiency. This depends on the nascent peptide, and a UAG stop codon contained in a stable stem-loop just past the 5 GGA codon. If you destabilize the loop, no bypass of intervening codons. If you put in compensatory nucleotides to create a new loop, bypass again. Putting GGA into 50 nts drops efficiency. Increasing 50 nt distance drops efficiency. Change GGAs to GCAs, still works although a little less bypass. Amino acids coded by upstream nucleotides 17-42 are crucial, since 1 base insertion or deletion to knock out of frame