[The construction of genomic library of Alcaligenes faecalis and cloning of nifH gene sequence homologous to Klebsiella pneumoniae].
A genomic library of Alcaligenes faecalis A-15Hl which possesses rather high nitrogenase activity has been constructed. The total DNA of A. faecalis A-15 Hl was partially digested with Sau3AI. 13-20 kb of fragments recovered from agarose gel were cloned in bacteriophage EM-BL4 vector. A total number of 1.2 x 10(6) of recombinants was obtained. It is much beyond the desired capacity of a library. By using nifH gene of K. pneumoniae from plasmid pGB1 as probe, we have successfully screened the clone containing its homologous sequence. The recombinant bacteriophage DNA was digested with EcoRI. A 3.5 kb of hybridizing band appeared after southern blotting and then was cloned in pUC19 DNA. The result of southern blotting indicated that the recombinant plasmid clone contained nifH gene sequence of A. faecalis. This clone was named as pAFH.
Hai
W
W
Institute for Application of Atomic Energy, CAAS.
Zheng
H
H
You
C
C
Wang
B
B
D000421:Alcaligenes / Q000235:genetics*; D005798:Genes, Bacterial; D015698:Genomic Library; D007711:Klebsiella pneumoniae / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid