Experimental manipulation of gene expression / edited by Masayori Inouye.

Date:
1983
Catalogue details

Licence: Attribution-NonCommercial 4.0 International (CC BY-NC 4.0)

Credit: Experimental manipulation of gene expression / edited by Masayori Inouye. Source: Wellcome Collection.

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    1. Use of Phage \ Regulatory Signals in E. cou 7 N N + 32°C 42°C 32°C 42°C Fig. 4 Coomassie Blue-stained SDS-polyacrylamide gel analysis of total cellular protein obtained from an N + - and an N ~ -defective lysogen carrying the plasmid рКСЗО^а/ЛГ. Protein samples were prepared from cells before (at 32°C) and 60 min after thermal induction (at 42°C). The position of the galactokinase gene product is indicated by an arrow. (M. Gribskow and R. Burgess, personal communication), and uvrA (Yoakum et al., 1982) from E. coli as well as certain Bacillus subtilis phage ф29 proteins (Garcia et al., 1982). In fact, the potential exists for any gene to be expressed that is preceded by a ribosome-binding signal recognized in E. coli. The gene is simply inserted on a blunt-ended DNA fragment into the blunt-ended Hpa\ site on рКСЗО. The appropriate DNA fragment can be generated (1) directly by restriction, (2) subsequent to the removal of single-strand overhanging ends by the action of Sj or mung bean nuclease, or (3) subsequent to fill-in of single-