Cloning and Expressing of Beefy Meaty Peptide Gene
| Autor: | Cheng-Wei Lin, 林錚威 |
|---|---|
| Rok vydání: | 2002 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 90 In this study, DNA recombinant technique was performed to construct a prokaryotic expression vector. The recombinant vector confers the transformed host cell the ability to produce beef meaty peptide. In this experiment, two complementary polynucleotides encoding beef meaty peptide were synthesized. The double strand polynucleotides with Sal 1 sticky end were inserted in pEZZ18, and then the recombinant plasmid（pEZZ18-BMP）were transformed to E.coli JM105. Some transformed E.coli colonies with the ability to produced umami peptide were isolated. The transformed E.coli were cultivated in LB broth, and the culture contained some umami peptide. The umami peptide linked with ZZ protein was isolated with an affinity chromatography. The isolated fusion protein was treated with CNBr to release the umami peptide. The umami peptide was purified with an HPLC system. The amino acid composition of the peptide was determined as those extracted from beef. The amino acid sequence was identified as that originally determined by Yamasaki and Maekawa. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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