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[Establishment of transgenic Xenopus laevis by intracytoplasmic sperm injection]
Pei-Zhou Jiang1, Xiang-Ling Feng, Hua Huang
1Cancer Research Institute, First Military Medical University, Guangzhou 510515, China.
Objective:
To study the feasibility of establishing transgenic laevis by intracytoplasmic sperm injection (ICSI).
Methods:
The testes of mature Xenopus laevis were taken for the purification of their sperms, which was subsequently incubated with digitonin to prepare concentrate of the sperms. Treatment of the concentrate with linearized reporter vector pCMV-EGFP-N1 was performed, and the sperms were then injected into unfertilized ova harvested from female laevis, followed by culture and observation of the development of the ova.
Results:
The condensed sperm we obtained were of high quality and after intracytoplasmic injection into the ova, a fertilization rate of 10% was achieved and 20% of the zygotes survived the neurula stages and developed into tadpoles, but all of which were slightly deformed. The integration ratio of green fluorescent protein (GFP) reporter gene was 81%, but GFP expression was not observed in the laevis.
Conclusion:
ICSI is a simple and practicable method for establishing transgenic Xenopus laevis.