Golden J W, Carrasco C D, Mulligan M E, Schneider G J, Haselkorn R
Department of Biology, Texas A&M University, College Station 77843.
J Bacteriol. 1988 Nov;170(11):5034-41. doi: 10.1128/jb.170.11.5034-5041.1988.
The filamentous cyanobacterium Anabaena sp. strain PCC 7120 produces terminally differentiated heterocysts in response to a lack of combined nitrogen. Heterocysts are found approximately every 10th cell along the filament and are morphologically and biochemically specialized for nitrogen fixation. At least two DNA rearrangements occur during heterocyst differentiation in Anabaena sp. strain PCC 7120, both the result of developmentally regulated site-specific recombination. The first is an 11-kilobase-pair (kb) deletion from within the 3' end of the nifD gene. The second rearrangement occurs near the nifS gene but has not been completely characterized. The DNA sequences found at the recombination sites for each of the two rearrangements show no similarity to each other. To determine the topology of the rearrangement near the nifS gene, cosmid libraries of vegetative-cell genomic DNA were constructed and used to clone the region of the chromosome involved in the rearrangement. Cosmid clones which spanned the DNA separating the two recombination sites that define the ends of the element were obtained. The restriction map of this region of the chromosome showed that the rearrangement was the deletion of a 55-kb DNA element from the heterocyst chromosome. The excised DNA was neither degraded nor amplified, and its function, if any, is unknown. The 55-kb element was not detectably transcribed in either vegetative cells or heterocysts. The deletion resulted in placement of the rbcLS operon about 10 kb from the nifS gene on the chromosome. Although the nifD 11-kb and nifS 55-kb rearrangements both occurred under normal aerobic heterocyst-inducing conditions, only the 55-kb excision occurred in argon-bubbled cultures, indicating that the two DNA rearrangements can be regulated differently.
丝状蓝细菌鱼腥藻Anabaena sp.菌株PCC 7120在缺乏化合态氮时会产生终末分化的异形胞。沿着藻丝大约每隔10个细胞就会发现一个异形胞,其在形态和生化方面都专门用于固氮。在鱼腥藻Anabaena sp.菌株PCC 7120异形胞分化过程中至少发生了两次DNA重排,这都是发育调控的位点特异性重组的结果。第一次是在nifD基因3'端内部发生11千碱基对(kb)的缺失。第二次重排在nifS基因附近发生,但尚未完全明确其特征。两次重排的重组位点处发现的DNA序列彼此没有相似性。为了确定nifS基因附近重排的拓扑结构,构建了营养细胞基因组DNA的黏粒文库,并用于克隆参与重排的染色体区域。获得了跨越界定该元件两端的两个重组位点之间DNA的黏粒克隆。该染色体区域的限制性图谱表明,重排是异形胞染色体上一个55-kb DNA元件的缺失。切除的DNA既未降解也未扩增,其功能(如果有)未知。55-kb元件在营养细胞或异形胞中均未检测到转录。该缺失导致rbcLS操纵子在染色体上位于距nifS基因约10 kb处。尽管nifD的11-kb重排和nifS的55-kb重排均在正常需氧异形胞诱导条件下发生,但只有55-kb的切除发生在氩气泡培养物中,这表明这两次DNA重排的调控方式可能不同。