AI Article Synopsis
- A genomic library was created from Alcaligenes faecalis A-15Hl, a strain with high nitrogenase activity, by partially digesting its DNA and cloning fragments into a bacteriophage vector, resulting in over 1.2 million recombinants.
- Using the nifH gene from Klebsiella pneumoniae as a probe, researchers screened the library and successfully identified a clone containing a homologous sequence.
- The cloned fragment, discovered through Southern blotting, was 3.5 kb and confirmed to contain the nifH gene sequence of A. faecalis, which was designated as pAFH.
Article Abstract
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.
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