Expression of GFP gene in mouse early embryo produced by sperm-mediated gene transfer.
- Author:
Huan-Sheng DONG
1
;
Yong-Wei FANG
;
Lan LI
;
Xiao-Lin LI
;
Li-Lan SUN
;
Xiang-Wei ZHAI
;
Qing-Jie PAN
;
Wei SHEN
Author Information
1. College of Animal Science and Technology, Institute of Animal Reproduction Development and Genetic Engineering, Laiyang Agricultural University, Qingdao 266109, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Embryo, Mammalian;
cytology;
metabolism;
Feasibility Studies;
Female;
Fertilization in Vitro;
Gene Expression;
Gene Transfer Techniques;
Green Fluorescent Proteins;
genetics;
metabolism;
Male;
Mice;
Mice, Inbred ICR;
Mice, Transgenic;
Microscopy, Fluorescence;
Oocytes;
cytology;
metabolism;
Spermatozoa;
cytology;
metabolism;
Zygote;
cytology;
metabolism
- From:
Chinese Journal of Biotechnology
2007;23(3):535-539
- CountryChina
- Language:Chinese
-
Abstract:
The efficiency of the exogenous DNA transfecting mouse sperm was studied by the DIG end labeled and immunohistochemistry technology. The results suggested that: the efficiency of transfecting positive rate of individual mouse sperm was distinct difference (P < 0.01), and the average rate was 13%. The acrosomal reaction was evaluated using the technology of Coomassie brilliant blue stained, and the appropriate in vitro fertilization (IVF) medium TYH was elected. Mouse sperms were transferred with GFP gene in vitro, and the mature oocytes were fertilized using IVF, and then the zygotes were cultured in vitro. The embryos were observed using the fluorescence microscopy, and the transgenic rate was 4.7%. The results suggested that sperm mediated gene transfer (SMGT) was an effective and feasible method.
