Peptides / Chinese Peptide Symposia (PDF)
Biology and Chemistry
(Sprache: Englisch)
The Fifth Chinese Peptide Symposium, hosted by Lanzhou University, was held at Lanzhou, China July 14-17, 1998, with 156 participants, including 30 scientists from abroad, representing nine countries. The four-day conference was both intense and spiritually...
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The Fifth Chinese Peptide Symposium, hosted by Lanzhou University, was held at Lanzhou, China July 14-17, 1998, with 156 participants, including 30 scientists from abroad, representing nine countries. The four-day conference was both intense and spiritually rewarding. Our goal for CPS-98 was to provide a forum for the exchange of knowledge, cooperation and friendship between the international and Chinese scientific communities, and we believe this goal was met. The symposium consisted of 8 sessions with 42 oral and 90 poster presentations, including synthetic methods, molecular diversity and peptide libraries, structure and conformation of peptides and proteins, bioactive peptides, peptide immunology, De Novo design and synthesis of proteins and peptides, ligand-receptor interactions, the chemistry-biology-interface and challenging problems in peptides. The enthusiastic cooperation and excellent contributions were gratifying and the active response of the invited speakers contributed to the success of the symposium. The presentations were of excellent caliber and represented the most current and significant aspects of peptide science. Dr. Kit Lam of the University of Arizona and Dr. Yun-Hua Ye of Peking University were the recipients of "The Cathay Award" sponsored by the H. H. Liu Education Foundation, offered for their seminal contribution in peptide science and the Chinese Peptide Symposium. Four outstanding young scientists were selected by the organizing committee to receive awards sponsored by Haikou Nanhai Pharmaceutical Industry Co. Ltd. (Zhong He Group).
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Detection of gene expression product of transgenic tobacco by antigenic peptide (p. 215-216) Jia-Xi Xu a*, Yuan Ma b, Mi Ma c and Zhong-Ping Lin d
aDepartment of Chemistry, Peking University. Beijing, 100871,
bDepartment of Chemistry, Tsinghua University, Beijing, 100084,
cInstitute of Botany, Academia Sinica, Beijing 10044,
dDepartment of Biology, Peking University, Beijing, 100871, China
Introduction
The need as well as the ability to cultivate special plants, such as antiviral wheat, antiinsect plants, economical plants etc, has increased with advances in biological science and technology and has created significant economic benefits. It is easy to detect DNA and RNA in transgenic processing. However, it is difficult to detect the protein as gene expression product due to low concentration and difficult separation and purification (1). Now we have used a synthetic antigenic peptide to solve this problem. After predicting and synthesizing an antigenic peptide of isopentyl transferase in transgenic tobacco, a key enzyme in gene expression, a peptide with the sequence IHARQQEQKF was conjugated to BSA. Its antibodies were then obtained from rabbit sera after immunizing the rabbit with this conjugated antigen. The antibody can be used for the qualitative and quantitative determination of gene expression product in crude proteinextract of leaf, stem and root of transgenic tobacco by ELISA and Western blot methods.
Results and Discussion
Prediction of epitope
The nucleotide and amino acid sequences of the isopentyl transferase in transgenic tobacco was obtained from published data (2). Its epitope was predicted according to the hydrophilicity (Hopp and Woods method), flexibility and accessibility by the computer program PC-Gene (3,4). Its secondary structure was predicted by the Chou and Fasman method (5). Peptide IHARQQEQKF (212-221 amino acid residues) with high hydrophilicity, flexibility and
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accessibility was predicted.
Synthesis of epitopic peptides
The predicted peptide was synthesized by Merrifield solid phase peptide synthesis method with the acid-labile tert-butyloxycarbonyl (Boc) group for temporary protection and acid-stable groups for side chain protection (3, 4, 6). Side chain-protected peptideresin was cleaved by anhydrous hydrogen fluoride under anisole, 1, 2-ethandithiol and thioanisole as scavengers and washed with cooled ethyl ether. Peptide was extracted with 30% acetic acid and purified by gel filtration on Sephadex G10. It was further purified by preparative HPLC and checked for homogeneity by reverse-phase HPLC. Its structure was confirmed by amino acid analysis and FAB-MS.
Immunologiccal methods
This antigenic peptide was conjugated to BSA by the glutaraldehyde method and its antibodies were then obtained from rabbit sera after immunizing rabbit with this conjugated antigen (7). The antibody was used to detect the gene expression product of transgenic tobaco at a dilution of 1:132 with OD492 nm 0.95 by the ELISA method. The results are shown in Table 1. This is a simple method for the qualitative and quantitative determination of gene expression product in crude protein-extract of leaf, stem and root of transgenic tobacco using ELISA and Western blot methods.
Synthesis of epitopic peptides
The predicted peptide was synthesized by Merrifield solid phase peptide synthesis method with the acid-labile tert-butyloxycarbonyl (Boc) group for temporary protection and acid-stable groups for side chain protection (3, 4, 6). Side chain-protected peptideresin was cleaved by anhydrous hydrogen fluoride under anisole, 1, 2-ethandithiol and thioanisole as scavengers and washed with cooled ethyl ether. Peptide was extracted with 30% acetic acid and purified by gel filtration on Sephadex G10. It was further purified by preparative HPLC and checked for homogeneity by reverse-phase HPLC. Its structure was confirmed by amino acid analysis and FAB-MS.
Immunologiccal methods
This antigenic peptide was conjugated to BSA by the glutaraldehyde method and its antibodies were then obtained from rabbit sera after immunizing rabbit with this conjugated antigen (7). The antibody was used to detect the gene expression product of transgenic tobaco at a dilution of 1:132 with OD492 nm 0.95 by the ELISA method. The results are shown in Table 1. This is a simple method for the qualitative and quantitative determination of gene expression product in crude protein-extract of leaf, stem and root of transgenic tobacco using ELISA and Western blot methods.
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Bibliographische Angaben
- 2006, 2002, 285 Seiten, Englisch
- Herausgegeben: Xiao-Yu Hu, Rui Wang, James P. Tam
- Verlag: Springer Netherlands
- ISBN-10: 0306468808
- ISBN-13: 9780306468803
- Erscheinungsdatum: 11.04.2006
Abhängig von Bildschirmgröße und eingestellter Schriftgröße kann die Seitenzahl auf Ihrem Lesegerät variieren.
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