Cloning and sequencing of the Pseudomonas aeruginosa 1244 pilin structural gene.
Cloning and sequencing of the Pseudomonas aeruginosa 1244 pilin structural gene.
复制标题
铜绿假单胞菌 1244 菌毛蛋白结构基因的克隆和测序。
DOI：
10.1007/bf00332233
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
Sadoff,JC
中科院分区：
文献类型：
--
作者：
Castric,PA;Sidberry,HF;Sadoff,JC
铜绿假单胞菌1244蛋白结构基因在cosmids和lambda中均有克隆。在p中检测到克隆基因的表达。利用单克隆抗体对铜绿杆菌PAO2003、PA103和653A进行免疫印迹检测。Western blot分析显示，克隆基因在菌株PAO2003和653A中表达的pilin略大于天然1244 pilin，而在菌株PA103中表达的pilin明显小于天然1244 pilin。1244菌毛特异性噬菌体不能裂解含有克隆的1244菌毛基因的菌株PAO2003，表明该重组菌株没有组装出功能性的1244菌毛。核苷酸测序显示一个编码区，当翻译时将产生一个15615道尔顿肽。该肽的氨基末端区域与已发表的pilin序列相同。虽然其余的肽通常不相同，但在潜在的抗原区域内可以看到共同的残基。
The pilin structural gene ofPseudomonas aeruginosa1244 was cloned in both cosmids and lambda. Expression of the cloned gene was detected inP. aeruginosastrains PAO2003, PA103, and 653A by an immunoblot reaction utilizing monoclonal antibodies. Western blot analysis showed that pilin expressed from the cloned gene was slightly larger than native 1244 pilin when produced in strains PAO2003 and 653A, but distinctly smaller in PA103. Bacteriophages specific for the 1244 pilus did not lyse strain PAO2003 containing the cloned 1244 pilin gene, indicating that functional 1244 pili were not assembled in this recombinant strain. Nucleotide sequencing revealed a coding region which when translated would produce a 15615 dalton peptide. The amino-terminal region of this peptide is identical with published pilin sequences. While the rest of the peptides are generally dissimilar, common residues are seen within potentially antigenic regions.