【作者】 梁姣； 王淑红； 张子平； 王艺磊；

【Author】 LIANG Jiao 1 , WANG Shu-hong 1 , ZHANG Zi-ping 2 ,WANG Yi-lei 1 ( 1 Fisheries College, Jimei University, Xiamen 361021, China; 2 Department of Biological Science, Seton Hall University, New Jersey, 07079, USA)

【机构】 集美大学水产学院； 西东大学生物系；

【摘要】 本研究建立了长距离实时定量(LA-real time)PCR技术检测DNA损伤的方法,并采用该方法检测了不同浓度的苯并(a)芘(Bap)对雄性斑马鱼(Danio rerio)不同组织的DNA损伤情况。实验采用腹腔注射Bap,浓度分别为1.0ng/g和10ng/g,二甲亚砜(DMSO)作为溶剂对照。注射24h、48h、96h后取脑,鳃,性腺和肌肉组织提取总DNA。采用LA-real time PCR技术扩增长度为14092bp的线粒体DNA,以237bp的短线粒体DNA片段作为参比片段。检测结果表明:与DMSO组相比,10ng/gBap组,24h斑马鱼鳃和肌肉组织,48h的鳃和脑组织以及1.0ng/g24h的鳃等组织的DNA均有显著损伤(p<0.05)。10ng/gBap组48h的精巢组织的DNA损伤程度极显著高于对照组(P<0.01)。本研究建立的长距离实时定量(LA-realtime)PCR技术可以用于检测Bap造成的DNA损伤。更多还原

【Abstract】 A novel long amplicon real-time (LA-real time) PCR was established to detect DNA damage, and the DNA damage of different tissues incuced by benzo (a) pyrene in male zebrafish (Danio rerio) also were studied by this method. The concentrations of BaP are 1.0ng/g and 10ng/g, dimethyl sulfoxide (DMSO) was used as solvent control. Chemicals were intraperitoneally injected, brain, gill, testis and muscle were dissected at 24h, 48h and 96h post injection. A 14092bp of mitochondrial DNA were amplificated from all tested control samples, and a 237bp short mitochondrial DNA was used as a reference fragment. The results show that, gill and muscle DNA at 24h post injection along with gill and brain DNA at 48h post injection from 10 ng/g Bap group, as well as gill DNA at 24h post injection from 1.0ng/g Bap group are significantly damaged comparing with that of DMSO group (p <0.05). Moreover, testis DNA at 48h post injection from 10 ng/g Bap group were highly significant damaged than that of solvent control group (p<0.01). The LA-real time PCR is capable of detecting DNA damage induced by Bap.更多还原