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首页> 外文期刊>Applied and Environmental Microbiology >Isolation and Characterization of Intracellular Protein Inclusions Produced by the Entomopathogenic BacteriumPhotorhabdus luminescens
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Isolation and Characterization of Intracellular Protein Inclusions Produced by the Entomopathogenic BacteriumPhotorhabdus luminescens

机译：病原细菌产生的细胞内蛋白质包裹体的分离和鉴定

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摘要

Cells of the entomopathogenic bacterium Photorhabdus luminescens contain two types of morphologically distinct crystalline inclusion proteins. The larger rectangular inclusion (type 1) and a smaller bipyramid-shaped inclusion (type 2) were purified from cell lysates by differential centrifugation and isopycnic density gradient centrifugation. Both structures are composed of protein and are readily soluble at pH 11 and 4 in 1% sodium dodecyl sulfate (SDS) and in 8 M urea. Electrophoretic analysis reveals that each inclusion is composed of a single protein subunit with a molecular mass of 11,000 Da. The proteins differ in amino acid composition, protease digestion pattern, and immunological cross-reactivity. The protein inclusions are first visible in the cells at the time of late exponential growth. Western blot analyses showed that the proteins appeared in cells during mid- to late exponential growth. When at maximum size in stationary-phase cells, the proteins constitute 40% of the total cellular protein. The protein inclusions are not used during long-term starvation of the cells and were not toxic when injected into or fed toGalleria mellonella larvae.
机译：致病性细菌光致发光细菌的细胞包含两种类型在形态上不同的晶体包涵蛋白。通过差速离心和等密度密度梯度离心从细胞裂解物中纯化出较大的矩形夹杂物（1型）和较小的双金字塔形夹杂物（2型）。两种结构均由蛋白质组成，并易于在pH 11和4下溶于1％的十二烷基硫酸钠（SDS）和8 M尿素中。电泳分析表明，每个包涵体均由单个蛋白质亚基组成，分子量为11,000 Da。这些蛋白质的氨基酸组成，蛋白酶消化模式和免疫学交叉反应性不同。在晚期指数生长时，蛋白质内含物首先在细胞中可见。蛋白质印迹分析表明，蛋白质在指数生长的中晚期出现在细胞中。当在固定相细胞中达到最大大小时，蛋白质占总细胞蛋白质的40％。蛋白质内含物在长期饥饿的细胞中不使用，并且当注射入或喂入家禽幼虫中时没有毒性。

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