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为探索人工诱导斑马鱼雄核发育二倍体克隆鱼的途径和方法,运用紫外线照射卵子使其遗传物质失活,利用斑马鱼体表花纹形态作为遗传标记,使用热休克技术阻止受精卵的第一次卵裂加倍染色体。试验结果表明总辐射剂量为120mJ cm2紫外线照射斑马鱼卵可以使雌核遗传物质完全失活;体表花纹为豹斑花纹被证实是由纯合隐性基因控制的性状,可以作为雄核发育遗传标记;遗传失活卵受精后经2min,(41 5±0 5)℃的热休克后出现两个染色体加倍的高峰期,分别为受精后的11min和23min,且后一时期效果优于前一时期,最高二倍体诱导率达46 15%;经遗传分析,共获得正常存活的雄核发育二倍体克隆鱼19尾,说明经过优化的紫外线结合热休克等染色体操作技术,可以成为诱导雄核发育二倍体克隆鱼的有效手段。
In order to explore ways and methods of artificially inducing diploid cloned fish in zebrafish male sterility, ultraviolet radiation was used to inactivate the genetic material of the diploid cloned fish. The zebrafish body surface morphology was used as a genetic marker, and the heat shock technique was used to prevent the A cleavage double chromosomes. The results showed that irradiation of zebrafish eggs with a total radiation dose of 120 mJ cm2 could completely inactivate the female genetic material. The surface pattern of the leopard pattern was confirmed to be controlled by homozygous recessive genes, After two days of heat shock at (41 5 ± 0 5) ℃, genetic inactivation eggs showed two chromosome doubling peaks after fertilization for 11 and 23 min, respectively, and the latter period was better than the former , The highest diploid inductivity was 46.15% .Through genetic analysis, 19 normal diploid cloned fish with normal survival were obtained, which indicated that the optimal UV-combined chromosome manipulation technique such as heat shock could be the inducing male Nuclear development of diploid cloned fish effective means.