Sokolov N N, El'darov M A, Anikeĭcheva N V, Karpychev I V, Samko O T, Fitsner A B, Kalugin A A, Khoroshutina E B, Skriabin K G
Bioorg Khim. 1991 Sep;17(9):1188-92.
A new site-specific endonuclease was detected in toluene lysates of Bacillus coagulans AUCM B-732 and designated as BcoAI. The enzyme was purified by fractionation of the cell-free extract in the two-phase PEG/dextran system followed by chromatography on DEAE-sepharose and phosphocellulose and shown to be free of nonspecific nucleases and phosphatases. BcoAI has three cleavage sites on lambda DNA, but does not cleave SV40, pBR322 and pUC19 DNA. BcoAI recognizes the sequence 5' CAC decreases GTG 3' on double-stranded DNA and cleaves it as indicated by the arrow to yield blunt-ended DNA fragments. Thus, BcoAI is a true isoschizomer of PmaCI from Pseudomonas maltophila C.
在凝结芽孢杆菌AUCM B - 732的甲苯裂解物中检测到一种新的位点特异性内切核酸酶,并将其命名为BcoAI。通过在双相聚乙二醇/葡聚糖系统中对无细胞提取物进行分级分离,然后在DEAE - 琼脂糖和磷酸纤维素上进行色谱分离来纯化该酶,结果表明其不含非特异性核酸酶和磷酸酶。BcoAI在λDNA上有三个切割位点,但不切割SV40、pBR322和pUC19 DNA。BcoAI识别双链DNA上的序列5' CAC↓GTG 3',并如箭头所示进行切割,产生平端DNA片段。因此,BcoAI是来自嗜麦芽假单胞菌C的PmaCI的真正同裂酶。