变铅青链霉菌dna磷硫酰化优先修饰区的功能分析-functional analysis of phosphothioacylate preferential modification region of streptomyces lividans dna.docxVIP

modification sequence.Rabbit polyclonal antibody against PNPase was raised using recombinant protein. Supershift assay confirmed that PNPase was one of the three shift bands.PNPase was known to function in RNA metabolism, such as mRNA degradation, RNA 3’-tail polymerization, etc. Its biological function of specific binding to preferential DNA phosphorothioate modification sequence remains to be elucidated.pnpase was reported to be essential in S.coelicolor. We are using overexpression and antisense RNA gene silencing techniques to investigate its biological function. Further study on PNPase will provide insight into the function of DNA preferential phosphorothioation.Keywords: phosphorothioate, Streptomyces lividans, pIJ101, preferential modification, fluorescent gel shift (EMSA), Supershift, polynucleotide phosphorylase (PNPase)符号说明Abbreviationsaac(3)IV Amp Aprbla bp BSAcml DMSO EBE.coli EDTA FPLC HEPESapramycin resistance gene ampicillinapramycinβ-lactamase gene base pairbovine serum albumin chloramphenicol acetyltransferase dimethyl sulfoxideethidium bromideEscherichia coliethylene diaminetetraacetic acid Fast Protein liquid chromatography4-(2-Hydroxyethyl)piperazine-1-ethanesulfon阿泊拉霉素抗性基因 氨苄青霉素 阿泊拉霉素β-内酰胺酶基因 碱基对 牛血清白蛋白 氯霉素酰基转移酶 二甲基亚砜 溴化乙啶大肠杆菌 乙二胺四乙酸 快速蛋白液相色谱4-羟乙基哌嗪乙磺酸kbic acid kilobases (pairs)千碱基对kDa km lacZ LC/MSMALDI- TOF-MSmin MSm/z nt OD ORFPFGE SDS TAEThio Tris tsrkilo-dalton kanamycinβ-galactosidaseliquid chromatography / mass spectrometry Matrix-assisted laser desorption /ionization time-of-flight mass spectrometryminutemass spectrometry ratio of mass/charge nucleotideoptical density open reading framepulsed field gel electrophoresis sodium dodecyl sulfateTris-acetate-EDTA buffer thiostrepton2-Amino-2-(hydroxymethyl)-1,3-propanediol thiostrepton resistance gene千道尔顿卡那霉素β-半乳糖苷酶 液相色谱质谱联用 基质辅助激光解析电离飞行 时间质谱分钟 质谱 质荷比 核苷酸光密度值 开放读码框 脉冲场凝胶电泳 十二烷基磺酸钠Tris-乙酸-EDTA 缓冲液 硫链丝菌素 三羟甲基氨基甲烷 硫链丝菌素抗性基因上海交通大学学位论文原创性声明本人郑重声明:所呈交的学位论文，是本人在导师的指导下， 独立进行研究工作所取得的成果。除文中已经注明引用的内容外，本