菌株ShigellaflexneriFB5响应氟磺胺草醚蛋白质组学分析.doc.docVIP

菌株Shigella flexneri?FB5响应氟磺胺草醚蛋白质组学分析1 1黑龙江省科技创新创业共享服务平台 哈尔滨市应用技术研究与开发项目（青年后备人才A类，编号：2015RAQYJ009） 摘 要：为明确氟磺胺草醚降解菌的蛋白质组学相关机理，针对实验室前期筛选得到的高效降解菌Shigella flexneri FB5，在明确菌株对氟磺胺草醚高效降解的基础上，本研究进行了蛋白提取和双向电泳试验，发现氟磺胺草醚诱导下菌株差异蛋白点13个，通过生物信息学比对得到其功能，根据氨基酸序列设计引物进行PCR扩增，得到氟磺胺草醚诱导下菌株两个相关基因，测序后发现基因F3序列与E.coli 结合蛋白dps基因同源性是100%，基因F6序列与沙门氏菌肠溶亚种血清变型索菲亚菌S1635外膜蛋白基因和E.coli SYW004外膜蛋白基因A相似度87%，同时对基因序列进行系统进化树构建，并且推测出基因的理化性质，为阐明该菌株降解机制奠定基础。 关键词：氟磺胺草醚；Shigella flexneri；双向电泳；PCR扩增 Strain Shigella flexneri FB5 response fomesafen proteomics analysis Abstract: To clear the proteomics mechanism of fomesafen degrading bacteria,?focusing on the highly efficient degrading bacterium Shigella flexneri FB5 screened and obtained in lab, based on the efficient degradation of fomesafen in this strain, protein extraction and two-dimensional electrophoresis were carried out and showed that the strain induced by fomesafen had 13paraproteins, and its function was obtained by bioinformatics alignment. Based on the amino acid sequence to design primers to be amplified by PCR. Two related genes of strain induced by fomesafen were gotten, after sequencing, it was found that the homology between the gene F3 sequence and the dps gene of E.coli-binding protein was 100%. The similarity of gene F6 sequence, salmonella enteric subtype serotype S1635 outer membrane protein gene and E.coli SYW004 external membrane protein gene A was 87%, at the same time carrying out systematical phylogenetic tree construction for the gene sequence, and speculating the physical and chemical properties of genes,which can be used to lay the foundation for clarification of the degradation mechanism of the strain. Key words:fomesafen; Shigella flexneri; two-dimensional electrophoresis; PCR amplification 1.引言 随着人们生活水平日益提高，工农业生产中除草剂的使用越来越广。除草剂的大量使用在给人们带来巨大利益收益和生活收益的同时，也会产生一些负面影响，如环境污染、生活在土壤中的微生物数量改变、杂草分群落生长、药害作用产生，以及杂草抗药性产生[1]。长残效除草剂不容易分解，短的存在时间可达1-3年，长的存在时间可超过4-6年[2-4]。氟磺胺草醚是一种二苯醚类除草剂，主要用于防治大豆、花生田产生的杂草，作用机制是抑制合成原卟啉原氧化酶，使敏感植物的细胞膜被破坏，最终导致细胞死亡