Liu Panpan, Zhou Yan, Wu Zhongqi, Zhong Hao, Wei Yanjun, Li Youzhi, Liu Shenkui, Zhang Yan, Fang Xuanjun
1Northeast Forestry University, Harbin, 150040 China.
2College of Life Sciences and Technology, Guangxi University, Guangxi, China.
3 Biotech. 2018 Jul;8(7):293. doi: 10.1007/s13205-018-1313-0. Epub 2018 Jun 26.
Mosquitocidal () strain S2160-1 was proposed to be an alternative to subsp. (). Discovering and validating a toxic gene by experimentation was a complex and time-consuming task, which can benefit from high-throughput sequencing analysis. In this research, we predicted and identified toxic proteins in the strain S2160-1 based on the draft whole genome sequence data. Through a local BLASP, 46 putative toxins were identified in S2160-1 genome, by searching against a customized toxin proteins database containing 653 protein or peptide sequences retrieved from public accessible resources and PCR/clone results in our laboratory ( value = 1e - 5). These putative toxins consist of 42 to 1216 amino acids. The molecular weights are ranged from 4.86 to 137.28 kDa. The isoelectric point of these candidate toxins varied from 4.3 to 10.06, and 16 out of which had a pH greater than 7.0. The analysis of tertiary structure and PFAM domain showed that 12 potential plasmid toxins may share higher similarity (9/12 QMEAN4 score > 0.3) with known toxins. In addition, functional annotation indicated that these 12 potential toxins were involved in "sporulation resulting in formation of a cellular spore" and "toxin activity". Moreover, multiple alignment and phylogenetic analysis were carried out to elucidate the evolutionary relationship among 101 known crystal or toxin proteins from public database and them with MEGA 6.0. It indicated that PS2160P2_1 and PS2160P2_153 may be potential Cry4-like toxins in S2160-1. This research may lay the foundation for future functional analysis of S2160-1 toxin proteins to reveal their biological roles.
杀蚊()菌株S2160 - 1被提议作为亚种()的替代品。通过实验发现和验证一个毒性基因是一项复杂且耗时的任务,这可以从高通量测序分析中受益。在本研究中,我们基于全基因组序列草图数据预测并鉴定了菌株S2160 - 1中的毒性蛋白。通过本地BLASTP,在S2160 - 1基因组中鉴定出46种假定毒素,通过与一个定制的毒素蛋白数据库进行比对,该数据库包含从公共可获取资源以及我们实验室的PCR/克隆结果中检索到的653个蛋白质或肽序列(期望值 = 1e - 5)。这些假定毒素由42至1216个氨基酸组成。分子量范围为4.86至137.28 kDa。这些候选毒素的等电点在4.3至10.06之间变化,其中16种的pH值大于7.0。三级结构和PFAM结构域分析表明,12种潜在的质粒毒素可能与已知的毒素具有更高的相似性(12种中有9种QMEAN4评分> 0.3)。此外,功能注释表明这12种潜在毒素参与“导致细胞孢子形成的孢子形成”和“毒素活性”。此外,进行了多序列比对和系统发育分析,以阐明来自公共数据库的101种已知晶体或毒素蛋白与它们之间的进化关系,使用的是MEGA 6.0。结果表明,PS2160P2_1和PS2160P2_153可能是S2160 - 1中潜在的Cry4样毒素。本研究可能为未来对S2160 - 1毒素蛋白进行功能分析以揭示其生物学作用奠定基础。