CLONING, SEQUENCING, AND MAPPING OF THE BACTERIOFERRITIN GENE (BFR) OF ESCHERICHIA-COLI K-12
CLONING, SEQUENCING, AND MAPPING OF THE BACTERIOFERRITIN GENE (BFR) OF ESCHERICHIA-COLI K-12
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DOI：
10.1128/jb.171.7.3940-3947.1989
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发表时间：
1989-07-01
影响因子：
3.2
通讯作者：
GUEST, JR
中科院分区：
文献类型：
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作者：
ANDREWS, SC;HARRISON, PM;GUEST, JR
大肠杆菌K-12的细菌铁蛋白(BFR)是一种储铁血球蛋白，以前被鉴定为细胞色素b1。细菌铁蛋白基因(BfR)已被克隆、测序，并定位于大肠杆菌连锁图谱中。最初是编码bfr-.lambda的基因融合。通过免疫筛选的方法检测到杂交蛋白(Mr 21,000)。含有大肠杆菌DNA Sau3A限制性片段的基因库。通过对含有bfR基因片段的大肠杆菌DNA的限制性内切酶进行Southern杂交，将bfR基因定位在大肠杆菌染色体物理图谱上的73min(str-SPC区)。然后从相应的lambda亚克隆完整的BfR基因。来自Kohara等人基因库的噬菌体。(Y.Kohara，K.Akiyama和K.Isono，Cell 50：495-508,1987)。BfR基因由474个碱基对和158个氨基酸密码子组成(包括起始密码子)，它编码的多肽与纯化的蛋白具有基本相同的大小(Mr 18495)和N-末端序列。在5‘’非编码区检测到一个潜在的启动子序列，但它与“铁盒”序列(即铁依赖的毛皮抑制蛋白的结合部位)无关。在含有BfR质粒的菌株中，BfR被扩增到总蛋白的14%。在BfR基因上游还检测到一个未知基因(Gen-)，它编码一个相对碱性的残基多肽，与BfR具有相同的极性。
The bacterioferritin (BFR) of Escherichia coli K-12 is an iron-storage hemoprotein, previously identified as cytochrome b1. The bacterioferritin gene (bfr) has been cloned, sequenced, and located in the E. coli linkage map. Initially a gene fusion encoding a BFR-.lambda. hybrid protein (Mr 21,000) was detected by immunoscreening a .lambda. gene bank containing Sau3A restriction fragments of E. coli DNA. The bfr gene was mapped to 73 min (the str-spc region) in the physical map of the E. coli chromosomes by probing Southern blots of restriction digests of E. coli DNA with a fragment of the bfr gene. The intact bfr gene was then subcloned from the corresponding .lambda. phage from the gene library of Kohara et al. (Y. Kohara, K. Akiyama, and K. Isono, Cell 50:495-508, 1987). The bfr gene comprises 474 base pairs and 158 amino acid codons (including the start codon), and it encodes a polypeptide having essentially the same size (Mr 18,495) and N-terminal sequence as the purified protein. A potential promoter sequences was detected in the 5'' noncoding region, but it was not associated with an "iron box" sequence (i.e., a binding site for the iron-dependent Fur repressor protein). BFR was amplified to 14% of the total protein in a bfr plasmid-containing strain. An additional unidentified gene (gen-64), encoding a relatively basic 64-residue polypeptide and having the same polarity as bfr, was detected upstream of the bfr gene.