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Alternative TitleFunctional studies of two outer membrane protein genes alr2269 and all3983 in Anabaena sp. PCC 7120
Thesis Advisor徐旭东
Degree Grantor中国科学院水生生物研究所
Place of Conferral水生生物研究所
Keyword鱼腥藻pcc7120 外膜蛋白 钙离子
Abstract鱼腥藻 PCC7120 (Anabaena sp. PCC7120) 是一种丝状固氮蓝藻,其细胞包被着质膜和外膜两层膜结构。本实验室曾鉴定了8个外膜蛋白,并初步观察到其中的两个外膜蛋白Alr2269 和All3983可能与钙离子的吸收利用相关。本研究以清楚的实验结果确证至少Alr2269是鱼腥藻在低浓度钙离子条件下生长所需要的。 主要研究内容和结果分3部分: 一、比较野生型与突变株alr2269::C.CE2、all3983::C.CE2的外膜蛋白证实,在突变株alr2269::C.CE2中All3983 表达上调,而突变株all3983::C.CE2的外膜蛋白与野生型无明显区别。 二、以luxAB为报告基因研究all3983的转录证实,在突变株alr2269::C.CE2中基因all3983 的转录受缺Ca2+诱导。all3983的转录水平在突变株alr2269::C.CE2中比在野生型中要高,与其蛋白质水平分析结果一致。 三、alr2269::C.CE2突变株在BG111/50ca中生长与野生型相比明显减缓,而导入野生型alr2269基因可恢复其野生型表型。all3983::C.CE2突变株在BG111/1000ca中才显示生长减缓,但互补未能恢复其野生型表型。 基于本研究结果,作者推测:Alr2269可能与钙离子的吸收利用有关。
Other AbstractAnabaena sp. PCC7120 is a filamentous nitrogen-fixing cyanobacterium whose cells are surrounded by an inner cytoplasmic membrane and an outer membrane. Eight outer membrane proteins were previously identified in our laboratory, and preliminary studies indicated that two such proteins, namely, Alr2269 and All3983, may be involved in calcium uptake and/or utilization. In this study, I present clear results to show that at least Alr2269 is required for the growth of Anabaena PCC7120 under low-calcium conditions. The main researches and results are described in three parts: 1. Comparison of Anabaena sp. PCC 7120 with mutant alr2269::C.CE2 or all3983::C.CE2 showed that All3983 was up-regulated in mutant alr2269::C.CE2, while no significant difference was found between the all3983::C.CE2 mutant and the wide type. 2. Reporter genes luxAB were used to analyze the transcription of all3983. The expression of all3983 in mutant alr2269::C.CE2 was induced upon calcium depletion. The transcription of all3983 in alr2269::C.CE2 was at a higher level than in the wide type, which was consistent with the 2-DE analysis at protein level. 3. The growth of the wide type and mutants on different calcium concentration was compared. The growth rate of alr2269::C.CE2 in BG111/50ca was significantly reduced in comparison to that of the wide type and restored to the wide type level after complementation with the wide type alr2269. In BG111/1000ca, mutant all3983::C.CE2 grew at a slightly lower rate than the wide type, but the complementation was not successful. Based on these results, it is hypothesized that Alr2269 is involved in calcium uptake and/or utilization.
Document Type学位论文
Recommended Citation
GB/T 7714
唐建新. 鱼腥藻PCC7120外膜蛋白基因alr2269和all3983的功能研究[D]. 水生生物研究所. 中国科学院水生生物研究所,2009.
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