摘要
采用反义转基因技术已成功获得整合CAsGnRHpc-antisense基因的转基因鲤鱼,简称AS-sGnRH F0鲤鱼。通过挤压1年龄AS-sGnRH F0鲤鱼的腹部,发现该群体中有精液/无精液个体比例为33/69,与对照组鲤鱼中有精液/无精液个体比例接近1:1相比存在显著差异。通过注射鲤鱼垂体抽提物诱导无精液AS-sGnRH F0鲤鱼恢复繁殖功能,获得3尾育性被恢复的AS-sGnRH F0鲤鱼。进一步的解剖观察发现,在无精液AS-sGnRH F0鲤鱼群体中,完全无性腺的AS-sGnRH F0鲤鱼比例约31.3%。根据直观的挤精液和解剖学观察,推测在AS-sGnRH F0鲤鱼群体中,雄鱼:雌鱼:“脂肪型”性腺个体的比例大约为35:45:20%。受精卵发育12 h后,统计A、B、C系AS-sGnRH F0鲤鱼精子的受精率分别为73.94、59.45和64.05%。从A、B、C系杂交后代F1分别随机取样100尾,PCR检测的阳性率分别为0、78.1和87.8%。研究结果表明通过补充外源激素可以使促性腺激素分泌不足而不育的AS-sGnRH F0鲤鱼恢复具有产生成熟配子的能力。
Transgenic carp expressed antisenese sGnRH mRNA were successfully cultivated by antisense transgene technology, which was abbreviated to AS-sGnRH F0 carp. By pressuring slightly the abdomen, it can be showed that the ratio of male carp was about 1/3 in one-year-old AS-sGnRH F0 group, which was much lower than that of control group. Three AS-sGnRH F0 carp were induced to produce mature mih from 69 AS-sGnRH F0 fish without semen by repeated intrapefitoneal injection of pituitary extract. By anatomical observation and assay, it was revealed that the ratio of carp without gonad was 31.3% in remanent 66 AS-sGmRH F0 carp without semen. So, it could be presumed that the proportion of ovary, spemlary and “fat” gonad individual was about 35, 45 and 20% in 102 AS-sGnRH F0 carp, respectively. 12 hours after fertilization, it was found that the fertilizing rate of sperm from A, B and C strain AS-sGnRH F0 was 73.94,59.45 and 64.05%, respectively. Then, 100 Juvenile fish was randomly sampied from hybrid F1 by mating AS-sGnRH F0 A, B and C strain semen with control ovum. By PCR assay, the rate of transgene fish with antisenese CAsGnRHpc gene was 0, 78.1 and 87.8%. The results showed that the fertility of hypogonadotrophic AS-sGnRH F0 carp by super-expressing antisense sGnRH RNA was able to be restored by renewing the extraneous hormone.
出处
《激光生物学报》
CAS
CSCD
2005年第5期348-352,共5页
Acta Laser Biology Sinica
基金
国家高技术研究发展计划(863计划)(2004AA213121)项目资助
关键词
转基因
反义RNA
鲤鱼
性腺发育
transgene
antisense RNA
common carp （ Cyprinus carpio）
gonad development
