Molecular cloning of nif DNA from Azotobacter vinelandii

Two clones which contained nif DNA were isolated from a clone bank of total EcoRI-digested Azotobacter vinelandii DNA. The clones carrying the recombinant plasmids were identified by use of the 32P-labeled 6.2-kilobase (kb) nif insert from pSA30 (which contains the Klebsiella pneumoniae nifK, nifD, and nifH genes) as a hybridization probe. Hybridization analysis with fragments derived from the nif insert of pSA30 showed that the 2.6-kb insert from one of the plasmids (pLB1) contains nifK whereas the 1.4-kb insert from the other plasmid (pLB3) contains nifD. Marker rescue tests using genetic transformation indicated that the 2.6-kb A. vinelandii nif fragment contains the wild-type alleles for the nif-6 and nif-38 mutations carried by Nif- strains UW6 and UW38. The 1.4-kb insert contains the wild-type allele for the nif-10 mutation carried by Nif- strain UW10.

[1]  C. Veeger,et al.  Advances in Nitrogen Fixation Research , 2011, Advances in Agricultural Biotechnology.

[2]  J. Ramos,et al.  Aspects of Genetics of Azotobacters , 1984 .

[3]  H. K. Das,et al.  Construction of a gene library from the nitrogen-fixing aerobe Azotobacter vinelandii. , 1983, Gene.

[4]  J. Vieira,et al.  The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. , 1982, Gene.

[5]  M. Norgard.  Rapid and simple removal of contaminating RNA from plasmid DNA without the use of RNase. , 1981, Analytical biochemistry.

[6]  P. Chambon,et al.  A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. , 1981, Analytical biochemistry.

[7]  G. Ruvkun,et al.  A general method for site-directed mutagenesis in prokaryotes , 1981, Nature.

[8]  P. Bishop,et al.  Evidence for an alternative nitrogen fixation system in Azotobacter vinelandii. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[9]  M. Summers,et al.  The bidirectional transfer of DNA and RNA to nitrocellulose or diazobenzyloxymethyl-paper. , 1980, Analytical biochemistry.

[10]  G. Ruvkun,et al.  Interspecies homology of nitrogenase genes. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[11]  K. Bott,et al.  Restriction enzyme analysis of Bacillus subtilis ribosomal ribonucleic acid genes , 1979, Journal of bacteriology.

[12]  R. Thayer,et al.  An improved method for detecting foreign DNA in plasmids of Escherichia coli. , 1979, Analytical biochemistry.

[13]  W. Page,et al.  Optimal conditions for transformation of Azotobacter vinelandii , 1979, Journal of bacteriology.

[14]  F. Ausubel,et al.  Physical map of chromosomal nitrogen fixation (nif) genes of Klebsiella pneumoniae. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[15]  F. Bolivar,et al.  Plasmids of Escherichia coli as cloning vectors. , 1979, Methods in enzymology.

[16]  W. Brill,et al.  Genetic analysis of Azotobacter vinelandii mutant strains unable to fix nitrogen , 1977, Journal of bacteriology.

[17]  K. Bott,et al.  Two-dimensional restriction analysis of the Bacillus subtilis genome: gene purification and ribosomal ribonucleic acid gene organization , 1977, Journal of bacteriology.

[18]  W. Brill,et al.  Transformation of nitrogen fixation genes in Azotobacter. , 1977, Basic life sciences.

[19]  W. Page,et al.  Physiological factors affecting transformation of Azotobacter vinelandii , 1976, Journal of bacteriology.

[20]  E. Southern.  Detection of specific sequences among DNA fragments separated by gel electrophoresis. , 1975, Journal of molecular biology.

[21]  T. Maniatis,et al.  Nucleotide sequence of the rightward operator of phage lambda. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[22]  W. Brill,et al.  Nitrogenase. 3. Nitrogenaseless mutants of Azotobacter vinelandii: activities, cross-reactions and EPR spectra. , 1973, Biochimica et biophysica acta.

[23]  H. Boyer,et al.  A complementation analysis of the restriction and modification of DNA in Escherichia coli. , 1969, Journal of molecular biology.

[24]  P. W. Wilson,et al.  Formation of the nitrogen-fixing enzyme system in Azotobacter vinelandii. , 1968, Canadian journal of microbiology.

[25]  D T Denhardt,et al.  A membrane-filter technique for the detection of complementary DNA. , 1966, Biochemical and biophysical research communications.

[26]  K. Miura,et al.  PREPARATION OF TRANSFORMING DEOXYRIBONUCLEIC ACID BY PHENOL TREATMENT. , 1963, Biochimica et biophysica acta.

[27]  P. W. Wilson,et al.  A Non-Gummy Chromogenic Strain of Azotobacter vinelandii , 1959, Nature.