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Articles by Harold G. Martinson
Total Records ( 2 ) for Harold G. Martinson
  Frank Rigo and Harold G. Martinson
  We have developed an in vitro transcription system, using HeLa nuclear extract, that supports not only efficient splicing of a multiexon transcript but also efficient cleavage and polyadenylation. In this system, both last-intron splicing and cleavage/polyadenylation are functionally coupled to transcription via the tether of nascent RNA that extends from the terminal exon to the transcribing polymerase downstream. Communication between the 3’ splice site and the poly(A) site across the terminal exon is established within minutes of their transcription, and multiple steps leading up to 3’-end processing of this exon can be distinguished. First, the 3’ splice site establishes connections to enhance 3’-end processing, while the nascent 3’-end processing apparatus makes reciprocal functional connections to enhance splicing. Then, commitment to poly(A) site cleavage itself occurs and the connections of the 3’-end processing apparatus to the transcribing polymerase are strengthened. Finally, the chemical steps in the processing of the terminal exon take place, beginning with poly(A) site cleavage, continuing with polyadenylation of the 3’ end, and then finishing with splicing of the last intron.
  Amer Jamil , Muhammad Yaqub , Munir A. Sheikh and Harold G. Martinson
  Cleavage and polyadenylation is an obligatory step in mRNA biogenesis in eukaryotes. Poly(A) tails play an important role in mRNA turn over, transport and translation. Currently available methods for the study of polyadenylation are very cumbersome and involve radioactivity. Dual luciferase assay was found to be an efficient and equally reliable method to CAT assay and RNase protection assay. Comparison of these methods is reported in this paper.
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