通过ELISA方法测定生长激素水平缩短转基因鱼的育种周期

【作者】 钟山； 汪亚平； 陈竹； 鲁鹏； 廖兰杰； 朱作言；

【机构】 中国科学院水生生物研究所淡水生态与生物技术国家重点实验室； 武汉大学生命科学学院遗传学系； 武汉大学基础医学院遗传学系；

【摘要】 为缩短快速生长转基因鱼育种年限并降低选育工作量,本研究采用了特研制的能检测鲤科鱼血清生长激素水平的单克隆抗体,结合酶联免疫吸附测定（enzyme-linked immunosorbentassays,ELISA）技术测定鲤鱼的血清生长激素水平,比较了快速生长转基因鲤鱼与普通鲤鱼杂交后F₄代中转基因鲤鱼与对照鲤鱼血清生长激素表达水平的差异,结果表明快速生长转基因鲤鱼F₄代其血清生长激素水平显著高于对照鲤鱼,转基因鲤鱼血清生长激素平均水平可为对照鲤鱼的52倍以上,并具有快速生长效应。结果提示在筛选快速生长转基因鱼亲本时,以往只选留转基因鱼,这种育种方式养殖成本高,并且后代筛选工作量大,而结合ELISA技术选留血清生长激素高的鱼育种则能降低养殖成本、降低后代筛选工作量并缩短育种的年限。更多还原

【Abstract】 In order to shorten the fast-growing transgenic fish breeding period and reduce the selection workload,this study employed a special monoclonal antibody that can detect the level of Cyprinidae fish’s serum growth hormone,and it is our unique approach.Fast-growing transgenic fish serum growth hormone level was analyzed by ELISA method and for the first time it applied the paired monoclonal antibodies.F₄-generation,the descent of fast-growing transgenic carp and common,were tested differently in their serum growth hormone between transgenic and control carps.The results show that the serum growth hormone level and growth rate of the fast-growing transgenic carp F₄-generations were significantly higher than the control carp’s.Transgenic carp was 52 times more than the control in average level of serum growth hormone with rapid growth effect.We also found that a simple detection by PCR always makes false positive outcomes,for some detected transgenic-positive fish had no rapid growth effect.Results point out that in the selection of fast-growing transgenic fish parents,only selection of transgenic fish will increase the cost and workload in the future generations,while monoclonal antibody technology was able to overcome the drawbacks and shorten the breeding period.更多还原