作　　者： ; ; ; ;

机构地区： 兰州医学院公共卫生学院

出　　处： 《癌变．畸变．突变》 2004年第3期144-147,F003,共5页

摘　　要： 背景与目的 :探索精子作为外源基因载体的可行性及阳离子脂质体的存在对精子转染率的影响。材料与方法 :取性成熟雄性金黄地鼠附睾内精子 ,采用共培养及脂质体介导两种方法与外源DNA共孵育后 ,用PCR、Southernblot及荧光原位杂交(FISH)等技术检测外源基因在精子细胞中的存在及存在部位 ,并对两种方法进行比较分析。结果 :金黄地鼠附睾内的活精子具有主动捕获外源基因的能力 ,荧光原位杂交证明部分外源基因进入了精子头部 ,脂质体介导组精子外源基因携带率较共培养组高 ,两组精子用DNA酶处理前后外源基因携带率间差异也有统计学意义 ,且观察发现死精子经洗涤后没有携带外源基因者。 结论 :即是在没有脂质体存在的情况下 ,活的精子细胞仍然能被外源基因所转染 ,这一过程涉及到精子对外源基因的主动性捕获 。 BACKGROUND&AIM: To explore the feasibility of sperm as foreign gene vectors and assess the effect of liposome during the sperm transfection. MATERIAL AND METHODS: Spermatozoa samples obtained from mature golden hamster epididymis were incubated with foreign DNA by co-culture and lipsome mediated methods.And two group samples were detected by PCR,Southern blot and FISH,and analyzed by statistics. RESULTS: Golden hamster spermatozoa could take exotic DNA voluntarily,further FISH research showed some DNA get into the head of the sperms.The transfection ratio of liposome mediated group was higher than co-culture group(P<0.001).There was significant difference in transfection ratio between two groups(P<0.05).No signals was found in dead spermatozoa after rinsed. CONCLUSION: Living spermatozoa can be transfected by foreign DNA even if liposome is absent,this transfection is most probably due to the active capture of the spermatozoa.Liposome can improve the foreign DNA transfection ratio of the spermatozoa.

关 键 词： 精子 基因载体 脂质体

领　　域： [生物学]