tailieunhanh - Báo cáo hóa học: " Comparative transcriptomic profile analysis of fed-batch cultures expressing different recombinant proteins in Escherichia coli"

Tuyển tập các báo cáo nghiên cứu về hóa học được đăng trên tạp chí hóa hoc quốc tế đề tài : Comparative transcriptomic profile analysis of fed-batch cultures expressing different recombinant proteins in Escherichia coli | Sharma et al. AMB Express 2011 1 33 http content 1 1 33 o AMB Express a SpringerOpen Journal ORIGINAL Open Access Comparative transcriptomic profile analysis of fed-batch cultures expressing different recombinant proteins in Escherichia coli Ashish K Sharma Shubhashree Mahalik Chaitali Ghosh Anuradha B Singh and Krishna J Mukherjee Abstract There is a need to elucidate the product specific features of the metabolic stress response of the host cell to the induction of recombinant protein synthesis. For this the method of choice is transcriptomic profiling which provides a better insight into the changes taking place in complex global metabolic networks. The transcriptomic profiles of three fed-batch cultures expressing different proteins viz. recombinant human interferon-beta rhIFN P Xylanase and Green Fluorescence Protein GFP were compared post induction. We observed a depression in the nutrient uptake and utilization pathways which was common for all the three expressed proteins. Thus glycerol transporters and genes involved in ATP synthesis as well as aerobic respiration were severely down-regulated. On the other hand the amino acid uptake and biosynthesis genes were significantly repressed only when soluble proteins were expressed under different promoters but not when the product was expressed as an inclusion body IB . High level expression under the T7 promoter rhIFN-b and xylanase triggered the cellular degradation machinery like the osmoprotectants proteases and mRNA degradation genes which were highly up-regulated while this trend was not true with GFP expression under the comparatively weaker ara promoter. The design of a better host platform for recombinant protein production thus needs to take into account the specific nature of the cellular response to protein expression. Keywords Transcriptomic profiling recombinant fed-batch Escherichia coli Introduction The wide variability in the expression levels of recombinant proteins in .