生物物理化学

プロテオミクスにおける基本的MS技術

試料作製および実際の質量分析法による解析

中西豊文

2002 年 46 巻 1 号 p. 1-6
発行日: 2002/03/15
公開日: 2009/03/31

DOIhttps://doi.org/10.2198/sbk.46.1

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The in-gel digestion followed by peptide mass fingerprint is an innovative technique of identification the proteins separated by two-dimensional polyacrylamide gel electrophoresis. It is widely used in various applications in proteomics, e.g., differential display of proteins in specific biological conditions, exhaustive identification of the components that physically interact in a protein complex. Accuracy from modern mass spectrometry instruments are typically in the 5 to 10ppm range which is in line with the search routine used in database mining and database identifications. In this chapter, the standardized procedures including sample preparation of vitreous humor, silver-staining, in-gel digestion, extraction of peptides from gel and mass spectrometric analysis are described.

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Phylogenetic analysis of feline calicivirus capsid gene in infected cats

佐藤良治, 大江香子, 福山正文, 古畑勝則, 岸川正剛, 鈴木嘉彦, 木内明男, 原元宣, 石川義久, 種野章

2002 年 46 巻 1 号 p. 7-14
発行日: 2002/03/15
公開日: 2009/03/31

DOIhttps://doi.org/10.2198/sbk.46.7

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To investigate the genetic diversity of feline calicivirus (FCV) in cases of field infection, we isolated FCV-S and FCV-B strains from 2 domestic cats (of 5 pet cats in the same house) that had an acute course and died, and an FCV-SAKURA strain from 1 of the remaining asymptomatic cats, and then RT-PCR-amplified part of the capsid region. The amplified cDNA products were cloned into TA cloning vectors, and the clones obtained were genetically analyzed. FCV-S and FCV-B yielded clones with same nucleotide sequences as FCV-SAKURA or containing somewhat different molecular species, with homologies ranging from 99.4% to 100%. On the other hand, FCV-SAKURA included clones with sequences common to FCV-S and FCV-B and, in addition, clones with different nucleotide sequences, with approximately 71.6% nucleotide homology. Phylogenetic analysis of these clones using 13 other Japanese isolates and 11 isolates from around the world showed that they formed 2 clearly different groups with a bootstrap value of 99.8%, suggesting that the asymptomatic cat was infected at least with a mixture of 2 FCV species with markedly different nucleotide sequences, and became super-infected with FCV from another cat which died.

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Polymorphism of glucose phosphate isomerase from mulberry (Morus spp.)

平野久, 村上理都子, 山ノ内宏昭, 江本泰二

2002 年 46 巻 1 号 p. 15-18
発行日: 2002/03/15
公開日: 2009/03/31

DOIhttps://doi.org/10.2198/sbk.46.15

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Glucose phosphate isomerase isoforms from leaves of 259 mulberry varieties were separated by starch gel electrophoresis. Varietal differences in the electrophoretic patterns of the isoforms were identified. A total of five patterns of the isoforms were detected in these varieties. The 259 varieties had specifically one of these patterns. The pattern similarity among varieties was not highly correlated to that of the other enzymes and proteins as previously reported, suggesting that mulberry varieties cannot be classified by the use of only a single enzyme or protein as a marker. However, it was found that the electrophoretic patterns can be used for variety identification with the aid of some characteristics such as the electrophoretic patterns of the other enzymes and proteins.

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