literaturereport生物专业原全英文文献汇报.pptxVIP

LiteratureReport生物专业原全英文文献汇报会计学Content1. High-level expression of the native barleyα-amylase/subtilisin inhibitor in Pichia pastoris2. A Manual for the Preparation and Transformation of Pichia pastoris Spheroplasts .High-level expression of the native barleyα-amylase/subtilisin inhibitor in Pichia pastorisContentIntroduction / ProgressMaterials and methodsResults Discussion Back1.Introduction/ Progress(1)Previously most plant proteins expressed in P.pastoris resulted in low yields (Cereghino and Cregg, 2000).(2) BASI(the Hordeum vulgare α-amylase/subtilisin inhibitor ) has previously been expressed in P. pastoris , but yields were very low (B?nsager et al., 2003).(3)Pichia pastoris is one of the eukaryotic expression hosts that have been developed into a highly successful expression system (Macauley-Patrick et al., 2005). 1.Introduction/ Progress(4) However, within the last couple of years, plant proteins have successfully been expressed in P. pastoris at high levels (Fierenset al., 2004; Yoneyama et al., 2007; Pan et al., 2007).(5) In this study we present successful high-level expression of the Hordeum vulgare α-amylase/subtilisin inhibitor (BASI) in P.pastoris. (From Journal of Biotechnology , 2008)(6) In this paper a high-level expression system for the bifunctional α-amylase/subtilisin inhibitor from barley is presented.1.Introduction/ Progress(7) Compared to previous attempts to express BASI in P. pastoris (B?nsager et al., 2003), various conditions have beenoptimized in this study, including signal sequence, codon usage, transformation technique and growth conditions. Back2. Materials and methods2.1. Strains, culture media and growth conditions(1)P. pastoris(Invitrogen, USA) GS115 was used for heterologous expression of BASI.(2) YPD medium (1% (w/v) yeast extract, 2% (w/v) peptone, 2% (w/v) dextrose) YNB (Yeast Nitrogen Base with ammonium sulphate without amino acids) BMSY media2. Materials and methods(3) To induce expression P. pastoris was grown