pMH2, a small plasmid bearing the nif gene cluster of Enterobacter agglomerans 333 as an excisable cassette.
A small plasmid containing the entire nif gene cluster of Enterobacter agglomerans 333 as an excisable cassette has been constructed, using pACYC177 as a vector. Two cosmid clones taken from a gene library of E. agglomerans plasmid pEA3 were used as a source of nif genes. A SmaI fragment of peaMS2-2, containing the H,D,K,Y,E,N,X,U,S,V,W,Z,M,L,A and B genes and an ApaI fragment of peaMS2-16 containing nif A,B,Q,F and J were selected to construct pMH2. The resulting plasmid of 33 kb carries the complete nif gene cluster as a nif cassette on a single XbaI fragment. The nif construct pMH2 in Escherichia coli strains has significant nitrogenase activity compared to wild-type E. agglomerans 333. The nif gene cluster construct was found to be very stable.
Stumpf
F
F
Lehrstuhl für Genetik, Universität Bayreuth, Germany.
Halda
L
L
Klingmüller
W
W
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004754:Enterobacter / Q000235:genetics*; D004926:Escherichia coli; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D005810:Multigene Family; D009586:Nitrogen Fixation / Q000235:genetics*; D010957:Plasmids; D011994:Recombinant Proteins
D004269:DNA, Bacterial; D011994:Recombinant Proteins