Establishment of transgenic Xenopus laevis by intracytoplasmic sperm injection

Pei Zhou Jiang, Xiang Ling Feng, Hua Huang, Xin Ming Shen, Jian Ling Shi, Kai Tai Yao

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Abstract

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.

Original languageEnglish (US)
Pages (from-to)673-677
Number of pages5
JournalDi 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA
Volume22
Issue number8
StatePublished - Aug 2002

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ASJC Scopus subject areas

  • Medicine(all)

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