小鼠卵母细胞及体细胞核移植重构早期端粒酶活性的测定  

Determination of the telomerase activity of mouse oocytes and early reconstructed embryos

作　　者：刘宇 张日欣 李钟淑[1] LIU YU, ZHANG Ri-xin, LI Zhong-shu(Animal Science Department,Agricultural College of Yanbian University ,Yanji ,Jilin 133000,Chin)

机构地区：[1]延边大学农学院动物科学系

出　　处：《中国兽医学报》2018年第3期586-590,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(413010028)

摘　　要：采用端粒酶重复序列扩增（telomeric repeat amplification protocol,TRAP）-银染法,对小鼠卵母细胞、体内受精胚胎及重构胚的各阶段端粒酶活性进行测定,对PCR产物的电泳条带进行灰度值分析,计算端粒酶总产品总量。结果表明：（1）体内受精胚胎及核移植重构胚各阶段的端粒酶活性随胚胎分裂呈上升趋势,囊胚期最高（P〈0.05）;（2）体内受精胚胎的端粒酶活性高于体细胞核移植重构胚端粒酶活性。同时进行囊胚计数并测定单卵裂球相对端粒酶活性,结果表明：单卵裂球的端粒酶活性呈下降趋势,囊胚期端粒酶活性最低（P〈0.05）。以上结果说明小鼠卵母细胞及体细胞核移植重构胚端粒酶活性变化与细胞发育水平及细胞发育全能性有关。Using telomeric repeat amplification protocol （TRAP）-silver staining,determined the te- lomerase activity of various stages of mouse oocytes,in vitro fertilized embryos and reconstructed embryos. Analyzed the electrophoresis of PCR products with gray value,calculated the total telomerase total product. The results showed that ： （1） telomerase activity of embryos and nuclear transfer reconstituted embryos increased with embryonic division,and the blastocyst stage reached the highest level （P〈0.05） ; （2） telomerase activity of in vivo fertilized embryos was higher than somatic cell nuclear transfer reconstructed embryos,Counted number of blastocyst and measured the relative telomerase activity of single blastomere, the results showed that the telomerase activity of single blastomeres were decreased and blastocyst stage reached the lowest level （P〈0.05）. The above results showed that the change of telomerase activity in mouse oocytes and somatic nuclear transfer reconstructed embryos were related to the level of cell development and cell developmen- tal totipotency.

关 键 词：核移植重构胚 端粒酶活性 TRAP-银染法 

分 类 号：S814.8[农业科学—畜牧学]