Cloning and expression of heterologous type IV fimbriae in pseudomonas aer

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Isfaqul Hussain
Isfaqul Hussain's picture
Cloning and expression of heterologous type IV fimbriae in pseudomonas aer

I want to clone and express type IV fimbriae of Dichelobacter nodosus in Pseudomonas aeruginosa for scaling up fimbrial antigen production. Please help me how can I proceed with commercially available suitable vectors. Is it fine to express in E. coli as well? 

Arvind Singh Pundir
Arvind Singh Pundir's picture
refer the material and

refer the material and methods section of this paper for reference
"Regulation of Type IV Fimbrial Biogenesis in Dichelobacter nodosus"
Material and methods :
Bacterial strains, plasmids, and media. Bacterial strains and plasmids are
listed in Table 1. Escherichia coli and P. aeruginosa strains were grown on 2X YT
medium (47) supplemented with 100 µg ampicillin/ml or 150 µg erythromycin/ml
for E. coli strains and 250 µg carbenicillin/ml for P. aeruginosa strains. D. nodosus
strains were grown on Eugon (Difco) yeast extract (EYE) agar containing 5%
defibrinated horse blood (Equicell) and supplemented with 1 µg erythromycin/
ml for selection of transformants or in Eugon (Difco) broth with yeast extract
in an anaerobic chamber as described previously (40).
Molecular techniques. Unless otherwise stated, molecular techniques were
performed as described elsewhere previously (40, 47). Chemically competent P.
aeruginosa cells were prepared as previously described (16) by using nutrient
yeast broth...........................................
jb.asm.org/cgi/reprint/188/13/4801.pdf

Isfaqul Hussain
Isfaqul Hussain's picture
Thanks for your kind reply.

Thanks for your kind reply. But I am a beginer, have not enough knowledge on cloning & expression. However, can anyone tell me about a comercial source of a vector containing lambda promoter with cI857 repressor that can be used in Ps. aeruginosa for expression of the fimbriae.