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Nucleic Acids Research, 1983, Vol. 11, No. 12 4241-4250
© 1983

MOLECULAR BIOLOGY

An open reading frame upstream from the nifH gene of Klebsiella pneumoniae

San-chiun Shen*, Zhong-tian Xue*, Qiu-tong Kong+ and Qi-long Wu+

*Division of Biology, California Institute of Technology Pasadena, CA 91125 +Laboratory of Molecular Genetics, Shanghai Institute of Plant Physiology Academia Sinica, Shanghai, China

Received March 21, 1983. Revised May 13, 1983. Accepted May 19, 1983.

An open reading frame upstream from nifHDK operon of Klebsiella pneumoniae had been described. The orientation of this open reading frame is opposite to that of nifHDK and sequence homology was found between the open reading frame promoter and the promoter of nifHDK operon.

A recombinant plasmid carrying the promoter region of the open reading frame fused to the ß-galactosidase gene was constructed. Strains of E.coli were transformed with the plasmid containing this open reading frame promoter-lacZ fusion or co-transformed with it and a plasmid carrying the nifA gene. An appreciable activity of ß-galactosidase was found in strains which received both plasmids, indicating that the promoter of the open reading frame can be activated by the product of nifA gene. Thus, the open reading frame found between nifHDK operon and nifJ behaves just like other nif genes of K.pneumoniae in requiring the product of nifA as the positive effector for expression.


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