[Establishment of transgenic Xenopus laevis by intracytoplasmic sperm injection].
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.


CONCLUSIONS
ICSI is a simple and practicable method for establishing transgenic Xenopus laevis.
Jiang
Pei-Zhou
PZ
Cancer Research Institute, First Military Medical University, Guangzhou 510515, China.
Feng
Xiang-Ling
XL
Huang
Hua
H
Shen
Xin-Ming
XM
Shi
Jian-Ling
JL
Yao
Kai-Tai
KT
D000818:Animals; D030801:Animals, Genetically Modified; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D008845:Microinjections; D020554:Sperm Injections, Intracytoplasmic / Q000379:methods*; D014982:Xenopus laevis / Q000235:genetics*
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins