Mulligan M E, Haselkorn R
Department of Molecular Genetics and Cell Biology, University of Chicago, Illinois 60637.
J Biol Chem. 1989 Nov 15;264(32):19200-7.
A second nitrogen fixation (nif) operon in the cyanobacterium (blue-green alga) Anabaena (Nostoc) sp. strain PCC 7120 has been identified and sequenced. It is located just upstream of the nifHDK operon and consists of four genes in the order nifB, fdxN, nifS, and nifU. The three nif genes were identified on the basis of their similarity with the corresponding genes from other diazotrophs. The fourth gene, fdxN, codes for a bacterial type ferredoxin (Mulligan, M. E., Buikema, W. J., and Haselkorn, R. (1988) J. Bacteriol. 167, 4406-4410). The four genes are probably transcribed as a single operon, but are expressed at a lower level than the nifHDK operon, and only after a developmentally induced DNA rearrangement occurs that excises a 55-kilobase pair element from within the fdxN gene (Golden, J. W., Mulligan, M. E., and Haselkorn, R. (1987) Nature 327, 526-529; Golden, J. W., Carrasco, C. D., Mulligan, M. E., Schneider, G. J., and Haselkorn, R. (1988) J. Bacteriol. 170, 5034-5041). The promoter for the nifB operon was located by primer extension. Comparison of the nifB 5'-flanking sequence with the nifH 5'-flanking sequence did not reveal any consensus base pairs that would define a nif promoter for Anabaena. The operon contains two instances of 7-base pair directly repeated sequences: seven copies of the repeated sequence are found between the nifB and fdxN genes and six copies are found between the nifS and nifU genes. The function of these repeats is unknown.
已在蓝细菌(蓝绿藻)鱼腥藻(念珠藻属)PCC 7120菌株中鉴定并测序了第二个固氮(nif)操纵子。它位于nifHDK操纵子的上游,由nifB、fdxN、nifS和nifU四个基因按此顺序组成。这三个nif基因是根据它们与其他固氮菌相应基因的相似性鉴定出来的。第四个基因fdxN编码一种细菌型铁氧化还原蛋白(穆利根,M.E.,布伊克马,W.J.,和哈塞尔科恩,R.(1988年)《细菌学杂志》167卷,4406 - 441页)。这四个基因可能作为一个单一操纵子进行转录,但表达水平低于nifHDK操纵子,并且只有在发生发育诱导的DNA重排后才会表达,这种重排会从fdxN基因内部切除一个55千碱基对的元件(戈尔登,J.W.,穆利根,M.E.,和哈塞尔科恩,R.(1987年)《自然》327卷,526 - 529页;戈尔登,J.W.,卡拉斯科,C.D.,穆利根,M.E.,施奈德,G.J.,和哈塞尔科恩,R.(1988年)《细菌学杂志》170卷,5034 - 5041页)。通过引物延伸确定了nifB操纵子的启动子。将nifB 5'侧翼序列与nifH 5'侧翼序列进行比较,未发现任何可定义鱼腥藻nif启动子的共有碱基对。该操纵子包含两个7碱基对的直接重复序列实例:在nifB和fdxN基因之间发现了七个重复序列拷贝,在nifS和nifU基因之间发现了六个重复序列拷贝。这些重复序列的功能尚不清楚。