多糖合成途径中功能基因的鉴定及其在粘细菌发育中的研究
关键词：粘细菌；子实体；冒险运动；社会运动；胞外基质多糖
Identification and developmental research of the polysaccharide synthesis genes from Myxococcus xanthus DK1622
Abstract： Myxobacteria are a group of prokaryotes that display complex multicellular behaviors. Their cell growth, predation, movement and development and other aspects show significantly social characteristics, which is currently known the only prokaryotes with multicellular behavior groups. Due to the characteristics of life special myxobacteria, it is considered as the most advanced prokaryotes. Because of the unique multicellular behavior and complex developmental regulatory mechanisms, it has been concerned by more and more researchers. Two different motility in fruiting body formation and the gliding motility (A motility, S motility) is the characteristic behavior of myxobacteria, in which intercellular signaling and extracellular matrix play an important role. Myxobacterial extracellular matrix is mainly composed of polysaccharides and proteins. The previous study found that bacteria cannot use monosaccharide and oligosaccharides, which presumably generate from amino acid by gluconeogenesis pathway. Given the important role of extracellular matrix in the mechanism of the development of the myxobacteria, it is of great significance to study the composition of extracellular matrix polysaccharides and the important functional genes in the process of the synthesis and metabolism of polysaccharides.
Key words: myxobacteria；fruiting body；adventurous motility；social motility；extracellular polysaccharide
目 录
摘要 1
关键词 1
Abstract. 1
Key words 1
引言 2
1 材料方法 3
1.1 菌株和质粒 3
1.2 引物 4
1.3 培养基与试剂 5
1.4 主要仪器 5
1.5 插入失活载体的构建 6
1.5.1 M. xanthus DK1622基因组的提取 7
1.5.2 多糖合成基因同源臂的扩增 7
1.5.3 PCR产物的T/A克隆 7
1.5.4 酶连产物的热激转化 8
1.5.5 质粒的提取 8
1.5.6 质粒的双酶切 8
1.5.7 琼脂糖凝胶DNA回收 8
1.5.8 pBJ113-同源臂载体的构建 8
1.6 粘细菌M. xanthus电转化 9
1.7 菌株插入失活的验证 9
1.7.1 DNA水平的验证 9
1.7.2 RNA水平的验证 9
1.8 序列分析软件与在线分析网址 11
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