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|Title:||The production of recombinant dengue virus E protein using Escherichia coli and Pichia pastoris||Authors:||Sugrue, R.J.
Cheong Chan, Y.
|Issue Date:||Dec-1997||Citation:||Sugrue, R.J., Cui, T., Xu, Q., Fu, J., Cheong Chan, Y. (1997-12). The production of recombinant dengue virus E protein using Escherichia coli and Pichia pastoris. Journal of Virological Methods 69 (1-2) : 159-169. ScholarBank@NUS Repository. https://doi.org/10.1016/S0166-0934(97)00151-1||Abstract:||The dengue virus envelope protein was expressed as a GST fusion protein using E. coli and P. pastoris as expression hosts. In E. coli the recombinant E protein is expressed initially as a soluble 81 kDa GST fusion protein. Treatment of the fusion protein with thrombin released a 55 kDa protein, which is the expected size for correctly processed, non-glycosylated recombinant E protein. The antiserum from animals immunised with this recombinant E protein was found to specifically recognise the dengue virus E protein in virus-infected cells, thus demonstrating the immunogenic nature of the recombinant E protein. This expression system allowed production of up to 2 mg of purified recombinant E protein from a 1 l bacterial culture. In contrast, expression of this GST fusion protein in P. pastoris is associated with extensive proteolytic degradation of the recombinant E protein. However, this proteolytic degradation was not observed in the truncated E protein sequences which were expressed. One of these recombinant fusion proteins, GST E401 was secreted into the culture medium at levels of up to 100 μg/l of growth medium.||Source Title:||Journal of Virological Methods||URI:||http://scholarbank.nus.edu.sg/handle/10635/112126||ISSN:||01660934||DOI:||10.1016/S0166-0934(97)00151-1|
|Appears in Collections:||Staff Publications|
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