Effect of combined treatment of caffeine benzamide and 137cs - ray on mutation frequency in soybean 射線復合處理提高大豆突變頻率的研究
An expression vector with fragment 9 in antisense orientation was constructed to block the expression of the relevant gene ( fragment 9 related gene , fnr gene ) in vero cells . interestingly , we found that the nontargeted mutation frequency induced by mnng was increased significantly , implicating that the product of the blocked gene may be involved in the inhibition of nontargeted mutation 利用反義核酸技術(shù)構(gòu)建含反向插入9號片段的真核細胞表達重組體并轉(zhuǎn)染細胞,以獲得反義rna阻斷vero細胞中相應基因的表達,發(fā)現(xiàn)mn ’ ng誘發(fā)的非定標性突變頻率顯著增高,提示被阻斷的相關(guān)基因的表達產(chǎn)物可能參與抑制非定標性突變的發(fā)生。
In our laboratory , a unique mutation detection system using a shuttle vector plasmid has been established to demonstrate that a low concentration of mnng ( 0 . 2 m ) can induce nontargeted mutation in mammalian cells : the mammalian cells were exposed to 0 . 2m mnng for 2 . 5h , then a shuttle plasmid pz189 carrying supf trna gene was transfected into cells after 24h culture . we found a 5 - fold higher mutation frequency of the plasmid replicated in pretreated cells than the spontaneous mutation frequency of the plasmid replicated in control cells . this kind of mutation did not occur immediately after mnng exposure 我們實驗室曾用一特殊的突變檢測系統(tǒng),直接證明dna損傷劑可在哺乳動物細胞誘發(fā)非定標性突變:首先用低濃度( 0 . 2 m )的短壽烷化劑mnng (半壽期為1 . 1hr )處理細胞2 . 5h后,繼續(xù)培養(yǎng)24h ,將重組有用作突變檢測的靶基因supftrna基因的穿梭質(zhì)粒pz189轉(zhuǎn)入細胞復制,發(fā)現(xiàn)在未受致癌物直接攻擊的穿梭質(zhì)粒中有較自發(fā)突變率高5倍以上的靶基因突變。