Rang Jie, He Hao, Wang Ting, Ding Xuezhi, Zuo Mingxing, Quan Meifang, Sun Yunjun, Yu Ziquan, Hu Shengbiao, Xia Liqiu
College of Life Science, Hunan Normal University, Hunan Provincial Key Laboratory of Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, Changsha, China.
PLoS One. 2015 Mar 17;10(3):e0119065. doi: 10.1371/journal.pone.0119065. eCollection 2015.
Bacillus thuringiensis is a widely used biopesticide that produced various insecticidal active substances during its life cycle. Separation and purification of numerous insecticide active substances have been difficult because of the relatively short half-life of such substances. On the other hand, substances can be synthetized at different times during development, so samples at different stages have to be studied, further complicating the analysis. A dual genomic and proteomic approach would enhance our ability to identify such substances, and particularily using mass spectrometry-based proteomic methods. The comparative analysis for genomic and proteomic data have showed that not all of the products deduced from the annotated genome could be identified among the proteomic data. For instance, genome annotation results showed that 39 coding sequences in the whole genome were related to insect pathogenicity, including five cry genes. However, Cry2Ab, Cry1Ia, Cytotoxin K, Bacteriocin, Exoenzyme C3 and Alveolysin could not be detected in the proteomic data obtained. The sporulation-related proteins were also compared analysis, results showed that the great majority sporulation-related proteins can be detected by mass spectrometry. This analysis revealed Spo0A~P, SigF, SigE(+), SigK(+) and SigG(+), all known to play an important role in the process of spore formation regulatory network, also were displayed in the proteomic data. Through the comparison of the two data sets, it was possible to infer that some genes were silenced or were expressed at very low levels. For instance, found that cry2Ab seems to lack a functional promoter while cry1Ia may not be expressed due to the presence of transposons. With this comparative study a relatively complete database can be constructed and used to transform hereditary material, thereby prompting the high expression of toxic proteins. A theoretical basis is provided for constructing highly virulent engineered bacteria and for promoting the application of proteogenomics in the life sciences.
苏云金芽孢杆菌是一种广泛使用的生物杀虫剂,在其生命周期中会产生多种杀虫活性物质。由于这些物质的半衰期相对较短,分离和纯化众多杀虫剂活性物质一直很困难。另一方面,这些物质在发育的不同阶段合成,因此必须研究不同阶段的样本,这进一步使分析复杂化。基因组学和蛋白质组学相结合的方法将提高我们识别此类物质的能力,特别是使用基于质谱的蛋白质组学方法。基因组和蛋白质组数据的比较分析表明,并非所有注释基因组推导的产物都能在蛋白质组数据中被识别。例如,基因组注释结果显示,全基因组中有39个编码序列与昆虫致病性相关,其中包括5个cry基因。然而,在获得的蛋白质组数据中未检测到Cry2Ab、Cry1Ia、细胞毒素K、细菌素、外切酶C3和肺泡溶素。还对芽孢形成相关蛋白进行了比较分析,结果表明绝大多数芽孢形成相关蛋白可以通过质谱检测到。该分析揭示了Spo0A~P、SigF、SigE(+)、SigK(+)和SigG(+),它们在孢子形成调控网络过程中都发挥着重要作用,也在蛋白质组数据中显示出来。通过对这两组数据的比较,可以推断出一些基因被沉默或表达水平很低。例如,发现cry2Ab似乎缺乏功能性启动子,而cry1Ia可能由于转座子的存在而不表达。通过这项比较研究,可以构建一个相对完整的数据库并用于转化遗传物质,从而促使毒性蛋白的高表达。为构建高毒力工程菌以及促进蛋白质基因组学在生命科学中的应用提供了理论依据。