兔受精卵显微注射外源基因后体外培养的卵裂发育

Cleavage in Vitro Culture of Rabbit Zygotes Microinjected oGH

从超数排卵的１４只母兔获得４３８枚受精卵，卵龄１６～２２小时．显微操作在带微分干涉和相差的Ｎｉｋｏｎ倒置显微镜下进行．注射针的尖端外也０．５μｍ，离尖端４０和８０μｍ处的外径分别为４．２和６．５μｍ．注射用外源基因是绵羊生长激素基因与ＭＴ－１启动基因藕连的线状ＤＮＡ溶液（１ｎｇ／μｌ）．１４０枚注射的受精卵和未注射的１４５枚受精卵（对照），在Ｈａｍ’ｓＦ—１０培养液（补充生长因子）中培养（３８℃，５％的ＣＯ２）．结果，培养４８小时后，注射组卵裂发育率分别是：未卵裂７．９％（１１／１４０）、卵裂至２～４细胞期１１．０％（１６／１４０）、卵裂至８～１６细胞期８０．７％（１１３／１４０）．对照组相应的卵裂率分别是４．１％（６／１４５）、１２．４％（１８／１４５）和８３．４％（１２１／１４５）．两组卵裂发育率相近．本实验的显微操作对注射后卵的发育没有产生明显的伤害影响．

zygotes collected from 14 superovulated rabbits were used in this study. Linear ovinegrowth hormone (oGH) gene combined with metallothonein gene promoter was microinjectedinto the male pronuclears under Nikon TMD - 2 invertal. ghcroseope. 140 injected and 145uninjected zygotes were cultured (38℃, 50% CO2 and humid air ) for 48 hours.In themicroinjected group, after the culture, of 140 zygotes, uncleavaged, cleavaged Minto 2-4 cell and8 - 16 cell were 7. 5%, 11. 0% and 80. 7% respectively. And in the uninjected group, thepercentage of corresponding to the microinjected group were 4. 1%, 18. 0% and 83. 4%respectively. The result showed that the microinjection manipulation in the study has no harm onthe cleavage of the injected rabbit zygotes.