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用精子载体法将MT-hGH基因导入泰山赤鳞鱼
翟玉梅1, 鹿培源1, 崔博文1, 王喆1, 时永香1, 杨晓梅1, 赵晶1, 岳永生2
1.山东大学生命科学学院 济南250100;2.山东农业大学动物科技学院 泰安271018
摘要:
于1998年3月–1999年2月，在山东农业大学采集泰山赤鳞鱼，采用精子载体法进行MT-hGH基因导入的研究。将泰山赤鳞鱼的精子在保存液内与含有MT-hGH基因的重组质粒保温30min，然后用于人工授精，共孵化出363尾幼鱼。待鱼苗长至5cm左右时，对其中的39尾进行PCR和Southern杂交检测。结果表明，有7尾鱼苗带有MT-hGH基因，转基因鱼的阳性率为17.95%。
关键词:  精子载体法  基因转移  泰山赤鳞鱼  MT-hGH基因
DOI：
分类号:
基金项目:山东省科学技术委员会攻关项目，D97060号
TRANSFERING MT-hGH GENE INTO VARICORIHINUS TRACROLEPIS WITH SPERM AS VECTORS
ZHAI Yu-mei1, LU Pei-yuan1, CUI Bo-wen1, WANG Zhe1, SHI Yong-xiang1, YANG Xiao-mei1, ZHAO Jing1, YUE Yong-sheng2
1.College of Life Sciences, Shandong University, Jinan, 250100;2.College of Animal Science, Shangdong Agriculture University, Taian, 271018
Abstract:
Transferring MT-hGH gene into Varicorihinus tracrolepis with sperm as vectors was carried out from March, 1998 to February, 1999. The Varicorihinus tracrolepis sperms were incubated with MT-hGH gene in dilute citrate solution for 30 minutes. Then they were used in artificial insemination. 997 eggs were inseminated with the treated sperms and 755 of them were fertilized. 363 fish were hatched in 3–4 days. When the fish grew to about 5cm, genomic DNA was isolated from 39 fish. The samples were tested by PCR and Southern blot hybridization. The result shows that 7 fish contained MT-hGH gene. The positive rate was about 17.95%.
Key words:  Sperm vectors, Gene transfer, Varicorihinus tracrolepis, MT-hGH gene
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