دورية أكاديمية
Expression and purification of recombinant tick anticoagulant peptide (Y1W/D10R) double mutant secreted by Saccharomyces cerevisiae.
| العنوان: | Expression and purification of recombinant tick anticoagulant peptide (Y1W/D10R) double mutant secreted by Saccharomyces cerevisiae. |
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| المؤلفون: | Cook JC; Department of Virus and Cell Biology, Merck Research Laboratories, West Point, Pennsylvania, 19486, USA., Schultz LD, Huang J, George HA, Herber WK, Ip C, Joyce JG, Mao SS, Markus HZ, Miller WJ, Sardana MK, Lehman ED |
| المصدر: | Protein expression and purification [Protein Expr Purif] 1998 Aug; Vol. 13 (3), pp. 291-300. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Academic Press Country of Publication: United States NLM ID: 9101496 Publication Model: Print Cited Medium: Print ISSN: 1046-5928 (Print) Linking ISSN: 10465928 NLM ISO Abbreviation: Protein Expr Purif Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Orlando, FL : Academic Press Original Publication: San Diego : Academic Press, c1990- |
| مواضيع طبية MeSH: | Peptides/*genetics , Saccharomyces cerevisiae/*genetics, Amino Acid Sequence ; Animals ; Arthropod Proteins ; Base Sequence ; Chromatography, High Pressure Liquid ; Chromatography, Ion Exchange ; Cloning, Molecular ; DNA, Recombinant ; Genetic Vectors ; Intercellular Signaling Peptides and Proteins ; Mating Factor ; Molecular Sequence Data ; Peptides/isolation & purification ; Recombinant Fusion Proteins/genetics ; Recombinant Fusion Proteins/isolation & purification ; Ticks/genetics |
| مستخلص: | A double mutant of tick anticoagulant peptide (TAP) was cloned as a chimeric fusion with the yeast alpha-mating factor pre-proleader peptide. Expression in yeast (Saccharomyces cerevisiae) resulted in the secretion of the TAP mutein into the culture medium. An HPLC-based assay was used to screen yeast strains to find those giving highest expression levels. Efficiency of cleavage at the junction of the leader-TAP mutein varied from strain to strain, and a rapid purification method followed by N-terminal sequence analysis was used to identify a host strain that minimized undesirable cleavage products. A purification scheme was developed which separated the TAP mutein from improperly processed peptides present in the medium. This scheme employed cation-exchange chromatography and reversed-phase HPLC. Scale-up of the process was successful and produced 100 mg of fully functional TAP mutein of >96% homogeneity from a 50-L yeast culture. (Copyright 1998 Academic Press.) |
| المشرفين على المادة: | 0 (Arthropod Proteins) 0 (DNA, Recombinant) 0 (Intercellular Signaling Peptides and Proteins) 0 (Peptides) 0 (Recombinant Fusion Proteins) 0 (tick anticoagulant peptide) 61194-02-3 (Mating Factor) |
| تواريخ الأحداث: | Date Created: 19980807 Date Completed: 19980917 Latest Revision: 20191210 |
| رمز التحديث: | 20221213 |
| DOI: | 10.1006/prep.1998.0893 |
| PMID: | 9693053 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1046-5928 |
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| DOI: | 10.1006/prep.1998.0893 |