EFFICIENT TRANSFORMATION OF PSEUDOMONAS STRAINS WITH PNI VECTORS BY ELECTROPORATION

被引:20
作者
ITOH, N [1 ]
KOUZAI, T [1 ]
KOIDE, Y [1 ]
机构
[1] AMANO PHARMACEUT CO LTD, RES LAB, NISHIKASUGAI, AICHI 481, JAPAN
关键词
D O I
10.1271/bbb.58.1306
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The optimum conditions for electro-transformation of some Pseudomonas strains including P. putida, P. fluorescens, and P. flavida, were defined using the pNI105 vector, resulting in 100-10,000 fold increases in transformation efficiency compared with conventional chemical transformation with MgCl2. The growth phase of the cultured cells and the field strength were important in obtaining high transformation efficiency. Under optimal conditions, 3.2 x 10(7) transformants per 1 mu g DNA were obtained using a combination of pNI105 and P. fluorescens IAM12022. This value is the highest ever obtained for a Pseudomanas species. In addition, the procedures were applicable to the effective transformation of fluorescent Pseudomonas strains with pNI vectors.
引用
收藏
页码:1306 / 1308
页数:3
相关论文
共 7 条
[1]  
DIVER JM, 1975, ANAL BIOCHEM, V189, P8
[2]   HIGH-EFFICIENCY TRANSFORMATION OF ESCHERICHIA-COLI BY HIGH-VOLTAGE ELECTROPORATION [J].
DOWER, WJ ;
MILLER, JF ;
RAGSDALE, CW .
NUCLEIC ACIDS RESEARCH, 1988, 16 (13) :6127-6145
[3]   TRANSFORMATION OF BACTERIA WITH PLASMID DNA BY ELECTROPORATION [J].
FIEDLER, S ;
WIRTH, R .
ANALYTICAL BIOCHEMISTRY, 1988, 170 (01) :38-44
[4]  
FRANKLIN FCH, 1985, DNA CLONING, V1, P165
[5]   NOVEL PLASMID VECTORS FOR GENE CLONING IN PSEUDOMONAS [J].
ITOH, N ;
KOIDE, Y ;
FUKUZAWA, H ;
HIROSE, S ;
INUKAI, T .
JOURNAL OF BIOCHEMISTRY, 1991, 110 (04) :614-621
[6]  
RODRIGUEZ RL, 1983, RECOMBINANT DNA TECH, P48
[7]   ELECTROPORATION AND EXPRESSION OF PLASMID PBR322 IN KLEBSIELLA-AEROGENES NCTC-418 AND PLASMID PRK2501 IN PSEUDOMONAS-PUTIDA CYM-318 [J].
TREVORS, JT .
JOURNAL OF BASIC MICROBIOLOGY, 1990, 30 (01) :57-61