氯霉素对集胞蓝细菌PCC 6803细胞中羧酶体数目及其相关蛋白丰度的影响
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国家自然科学基金项目(31570048);中央高校基本科研业务费专项(2014PY003)


Effects of chloramphenicol on carboxysome number and cellular abundance of CcmK2 in Synechocystis sp. PCC 6803
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    摘要:

    羧酶体是由致密的外壳蛋白包裹着核酮糖1,5二磷酸羧化酶/氧化酶和碳酸苷酶形成的细胞内蛋白质小体,是蓝细菌二氧化碳固定的场所。为解析羧酶体在不同培养条件下的功能,本研究构建了原核表达载体pET28aCcmK2,并在大肠杆菌BL21(DE3)中诱导表达带有组氨酸标签的CcmK2重组蛋白,制备了其多克隆抗体,探讨了不同质量浓度氯霉素对细胞内羧酶体数目和羧酶体相关蛋白丰度的影响。首先,确定了不同质量浓度的氯霉素对集胞蓝细菌PCC 6803生长的影响,发现5.0 μg/mL氯霉素能够完全抑制菌株的生长,但是可以维持菌株存活。随后,探讨了集胞蓝细菌在葡萄糖异养和光合自养转换过程中,氯霉素对细胞内羧酶体数目及羧酶体相关蛋白的影响。结果显示:氯霉素的加入抑制了细胞内羧酶体数目对环境碳源的响应;同时,Western blot检测发现5.0 μg/mL氯霉素能够抑制细胞内新的羧酶体外壳蛋白CcmK2的合成。这些结果表明,氯霉素可能通过抑制羧酶体外壳蛋白的合成来干扰细胞内羧酶体的形成,细胞内羧酶体对环境碳源的响应存在复杂的调控机制。

    Abstract:

    Cyanobacteria plays a significant role in the global carbon cycle.As the place for carbon dioxide fixation,carboxysome is a proteinaceous compartment that encapsulates the carbonfixing enzyme ribulose1,5 -bisphosphate carboxylase (RuBisCO) and carbonic anhydrase.In this study,the effects of chloramphenicol on carboxysome number and cellular abundance of CcmK2 in Synechocystis sp. PCC 6803 were analyzed.A batch of test were performed to study the inhibited effect of chloramphenicol on the growth of -Synechocystis- sp. PCC 6803 and its derivatives.Results showed that 5.0 μg/mL of chloramphenicol inhibited bacterial multiply completely.From heterotrophic status to photoautotrophic growth,the number of carboxysome rapidly increased in 12 h,but the addition of 5.0 μg/mL chloramphenicol inhibited effectively the increase of cellular carboxysome number.Result of Western blot analysis showed that the abundance of the most abundant carboxysome shell protein CcmK2 was inhibited by this antibiotics.It is indicated that chloramphenicol could inhibit novel carboxysome shell protein and disturb the carboxysome formation under different carbonenvironment.Further studies on carboxysome assembly are needed.

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李松柏,支瑶,李盼,汪方奎,陈雯莉.氯霉素对集胞蓝细菌PCC 6803细胞中羧酶体数目及其相关蛋白丰度的影响[J].华中农业大学学报,2018,37(1):7-11

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  • 收稿日期:2017-05-17
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  • 在线发布日期: 2018-01-02
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