mutation detection of abcd1 gene in the molecular diagnosis of x-linked adrenoleukodystrophy by denaturing hplc 法在腎上腺腦白質(zhì)營(yíng)養(yǎng)不良分子診斷中的應(yīng)用
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(shí)驗(yàn)室曾用一特殊的突變檢測(cè)系統(tǒng),直接證明dna損傷劑可在哺乳動(dòng)物細(xì)胞誘發(fā)非定標(biāo)性突變:首先用低濃度(0.2m)的短壽烷化劑mnng(半壽期為1.1hr)處理細(xì)胞2.5h后,繼續(xù)培養(yǎng)24h,將重組有用作突變檢測(cè)的靶基因supftrna基因的穿梭質(zhì)粒pz189轉(zhuǎn)入細(xì)胞復(fù)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。