Scott K F, Rolfe B G, Shine J
J Mol Appl Genet. 1981;1(1):71-81.
A DNA fragment carrying the Klebsiella pneumoniae nifK, D, and H genes was isolated from the nif- strain UNF841 (Tn5::nifK) by molecular cloning into the Escherichia coli plasmid pBR325. The nucleotide sequences of both the nifH gene, which encodes the Fe protein of the nitrogenase enzyme complex, and 622 nucleotides of the nifD gene, which encodes the alpha-subunit of the Mo-Fe protein, were determined by direct DNA sequencing by both the chemical and chain termination methods. A comparison of the primary structure of the Klebsiella nifH gene and its product with that recently determined for the blue-green alga Anabaena demonstrates that the gene sequences are more divergent than the protein sequence data would suggest. This implies that despite the strong, presumably functional, constraints that act at the protein structure level, the nucleotide sequence of the gene and its mRNA are only restrained by the coding requirements, allowing substantial drift in codon usage.
通过分子克隆到大肠杆菌质粒pBR325中,从固氮缺陷型菌株UNF841(Tn5::nifK)中分离出携带肺炎克雷伯菌nifK、D和H基因的DNA片段。通过化学法和链终止法直接进行DNA测序,确定了编码固氮酶复合物铁蛋白的nifH基因以及编码钼铁蛋白α亚基的nifD基因622个核苷酸的核苷酸序列。将肺炎克雷伯菌nifH基因及其产物的一级结构与最近测定的蓝藻鱼腥藻的结构进行比较,结果表明基因序列的差异比蛋白质序列数据所显示的更大。这意味着尽管在蛋白质结构水平上存在强大的、可能起作用的功能限制,但基因及其mRNA的核苷酸序列仅受编码要求的限制,从而允许密码子使用上有相当大的漂移。