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期刊论文
Cloning and Identif ication of promoter of Pseudomonas Pseudoaligenes①
HIGH TECHNOLOGY LETTERS Sep. 2003 Vol. 9 No.3,-0001,():
Promoter-probe vector p SUPV4 is used to clone the promoter of Pseudomonas pseudoal-caligenes directly in E. coli, and the recombinant p PA7, which has the highest kanamycin resis-tance, is obtained. Sequencing the PA7 fragment discloses several motifs similar to the conserva-tive domains of prokaryotic promoters, including-10box, -35box, parallel SD fragment es-sential to transcription initiation, and the translation initiation site ATG. Southern blotting of PA7 indicates that the PA7 fragment comes from P. pseudoalcali genes genome and has probably one copy. The PA7 fragment is subcloned by PCR, and the result shows that the 5'-flanking fragment from 889 to 1120 bp has promoter activity, which can be enhanced by the 0.7Kb f rag-ment at 5'-end. The fragments of p PA7 and p PA7-2 are transferred into pseudomonas pseudoali-genes by elect roporation, and the significant higher kanamycin resistance of transformants than that of control indicates that the PA7 fragment has the promoter activity in P. pseudoali gene.
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