相关基因pyrD的克隆与功能分析.PDFVIP

微生物学通报 JUL 20, 2010, 37(7): 981−985 Microbiology China © 2010 by Institute of Microbiology, CAS tongbao@ 研究报告 Pseudomonas pseudoalcaligenes B50 铁载体合成 相关基因pyrD 的克隆与功能分析 丁延芹 黄伟红 姚良同 于素芳 杜秉海* (山东农业大学生命科学学院微生物学系 山东 泰安 271018) 摘 要: 从棉花根际分离的一株细菌B50 在低铁条件下可以产生铁载体。通过形态学特征、生理 生化特征及其 16S rDNA 序列分析, 将其鉴定为Pseudomonas pseudoalcaligenes 。采用三亲本杂交 的方法将转座子 Tn5-1063a (含 luxAB) 导入 B50 中, 进行转座子插入诱变, 获得突变株。用 TAIL-PCR 方法得到与铁吸收有关的pyrD 基因序列。通过互补实验验证pyrD 与铁载体的合成有 关。Pseudomonas pseudoalcaligenes 能够产生铁载体属于首次报道。 关键词: Pseudomonas pseudoalcaligenes , 铁载体合成, pyrD Cloning and Functional Analysis of pyrD Gene Involved in Siderophores Biosynthesis in Pseudomonas pseudoalcaligenes B50 * DING Yan-Qin HUANG Wei-Hong YAO Liang-Tong YU Su-Fang DU Bing-Hai (Department of Microbiology , College of Life Sciences, Shandong Agricultural University, Taian, Shandong 271018, China) Abstract: In order to study the mechanism of bacteria producing siderophore, B50 was isolated from the cotton rhizosphere. The strain B50 can produce siderophore under iron-limiting conditions. It was identified as Pseudomonas pseudoalcaligenes based on its morphological, physical-chemical character- istics and 16S rDNA sequence. Mutants of B50 were obtained through conjugation. The pyrD gene was cloned from the mutant deficient in siderophores synthesis by TAIL-PCR (Thermal asymmetric inter- laced). The pyrD gene is required