Owttrim G W, Coleman J R
Department of Botany, University of Toronto, Ontario, Canada.
J Bacteriol. 1989 Oct;171(10):5713-9. doi: 10.1128/jb.171.10.5713-5719.1989.
The expression of the cyanobacterial recA gene, isolated from Anabaena variabilis, has been examined at the levels of transcript and protein abundance. Exposure of the cyanobacterium to a variety of DNA-damaging agents, including mitomycin C, methyl methanesulfonate, and UV irradiation, results in a rapid increase in the abundance of the recA transcript above basal levels as determined by Northern (RNA) blot analysis. A concomitant increase in the abundance of a 37- to 38-kilodalton polypeptide was also detected by Western (immuno-) blot analysis of soluble cyanobacterial polypeptides using polyclonal antiserum directed against the Escherichia coli recA protein. The cyanobacterial polypeptide is of the same molecular mass as that synthesized by an in vitro, DNA-directed procaryotic transcription-translation system primed with an A. variabilis genomic fragment containing the recA gene. Nucleotide sequence analysis of the cyanobacterial gene revealed a protein of 358 amino acids with a molecular weight of 38,403 daltons. The A. variabilis and E. coli recA genes share similarity at 58% of the amino acid residues; however, an E. coli-like lexA repressor-binding site is not present in the A. variabilis promoter region. The similarities of A. variabilis and E. coli recA expression and gene sequence are discussed.
对从多变鱼腥藻中分离出的蓝藻recA基因在转录本和蛋白质丰度水平上进行了检测。用Northern(RNA)印迹分析确定,将该蓝藻暴露于多种DNA损伤剂,包括丝裂霉素C、甲基磺酸甲酯和紫外线照射下,recA转录本的丰度会迅速增加,高于基础水平。使用针对大肠杆菌recA蛋白的多克隆抗血清对可溶性蓝藻多肽进行Western(免疫)印迹分析,也检测到一种37至38千道尔顿多肽的丰度随之增加。该蓝藻多肽的分子量与用含有recA基因的多变鱼腥藻基因组片段启动的体外DNA指导的原核转录-翻译系统合成的多肽相同。对该蓝藻基因的核苷酸序列分析揭示了一种由358个氨基酸组成、分子量为38403道尔顿的蛋白质。多变鱼腥藻和大肠杆菌的recA基因在58%的氨基酸残基上具有相似性;然而,多变鱼腥藻的启动子区域不存在类似大肠杆菌的lexA阻遏物结合位点。本文讨论了多变鱼腥藻和大肠杆菌recA表达及基因序列的相似性。