Nakano T, Okaichi K, Harada K, Matsumoto H, Kimura R, Yamamoto K, Akasaka S, Ohnishi T
PL Botanical Institute, PL Gakuen Women's Junior College, Osaka, Japan.
Mutat Res. 1995 Mar;336(2):153-9. doi: 10.1016/0921-8777(94)00053-9.
A shuttle vector, pZ189, carrying a bacterial suppressor tRNA marker gene (supF) was dissolved in Tris-EDTA buffer containing 0.3 M 10B-enriched boric acid and then irradiated with boron neutron captured beam (BNCB) produced by the nuclear reaction 10B (n,alpha) 7Li with thermal neutrons. A DNA repair-deficient mutant, KS46 (uvrA-), of Escherichia coli was transformed with the plasmid DNA, and the transformants carrying mutations on the supF gene were selected as nalidixic acid-resistant colonies. The mutation frequency (2.4 x 10(-4)) of pZ189 at the D10 dose was about 70 times greater than the spontaneous rate (3.5 x 10(-6)). The plasmid mutations were analyzed using DNA sequencers; 88% of them were base substitutions. A few minus-one frameshifts (7%) and deletions (5%) were detected. Among these base substitutions, transversions of G:C to T:A (42%) and G:C to C:G (29%) predominated. Twenty-seven percent of the base substitutions were G:C to A:T transitions; no A:T to G:C transitions were detected.
一个携带细菌抑制性tRNA标记基因(supF)的穿梭载体pZ189溶解于含有0.3M富集硼-10的硼酸的Tris-EDTA缓冲液中,然后用由热中子引发的10B(n,α)7Li核反应产生的硼中子俘获束(BNCB)进行辐照。用该质粒DNA转化大肠杆菌的DNA修复缺陷型突变体KS46(uvrA-),并将携带supF基因突变的转化体筛选为耐萘啶酸菌落。在D10剂量下,pZ189的突变频率(2.4×10(-4))比自发率(3.5×10(-6))高约70倍。使用DNA测序仪分析质粒突变;其中88%为碱基替换。检测到少数-1移码突变(7%)和缺失(5%)。在这些碱基替换中,G:C到T:A的颠换(42%)和G:C到C:G的颠换(29%)占主导。27%的碱基替换是G:C到A:T的转换;未检测到A:T到G:C的转换。