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https://hdl.handle.net/1822/33926
Título: | Cloning and expression of carbohydrate binding module in Pichia pastoris |
Autor(es): | Moreira, Susana Domingues, Lucília Gama, F. M. Casal, Margarida |
Data: | 2005 |
Citação: | Moreira, Susana; Domingues, Lucília; Casal, Margarida; Gama, F. M., Cloning and expression of carbohydrate binding module (CBM) in Pichia pastoris. CBM6 - 6th Carbohydrate Bioengineering Meeting. Barcelona, Spain, 3-6 April, 2005. |
Resumo(s): | The main goal of this work is the production of recombinant biologically active peptides fused with a Carbohydrate Binding Module (CBM). Aiming at the optimization of large scale expression, CBM peptide production was done by cloning CBM coding sequence in two different systems of Pichia pastoris: pGAPZC which has a constitutive promoter and pPICZC which has an inductive promoter. The integration of the CBM coding sequence, in yeast genome, was confirmed by slot-blot for both expression systems. Transcription was analysed by northern-blot and SDS-PAGE. The results obtained with these two expression systems were different. Apparently, there were no clones of P. pastoris transformed with pGAPZC-CBM that had produced any protein with starch affinity, under the batch and fed-batch conditions tested in this work. On the other hand, only one studied clone of P. pastoris transformed with pPICZC-CBM vector had produced, in batch conditions, a protein with affinity for starch. However, under fed-batch conditions, the results obtained with this clone were not conclusive, suggesting that conditions for large scale production must be optimized. |
Tipo: | Resumo em ata de conferência |
URI: | https://hdl.handle.net/1822/33926 |
Arbitragem científica: | yes |
Acesso: | Acesso aberto |
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Ficheiros deste registo:
Ficheiro | Descrição | Tamanho | Formato | |
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document_7928_1.pdf | 28,45 kB | Adobe PDF | Ver/Abrir |