Production of porcine cloned transgenic embryos expressing green fluorescent protein by somatic cell nuclear transfer

作     者：ZHANG Yunhai1, PAN Dengke1,2, SUN Xiuzhu1, SUN Guojie1, WANG Xiaobo1, LIU Xiaohui1, LI Yan1, DAI Yunping1 & LI Ning1 1. State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing 100094, China 2. College of Animal Science and Technology, China Agricultural University, Beijing 100094, China 

作者机构：State Key Laboratory for Agrobiotechnology China Agricultural University Beijing China College of Animal Science and Technology China Agricultural University Beijing China 

出 版 物：《Science China(Life Sciences)》 (中国科学（生命科学英文版）)

年 卷 期：2006年第49卷第2期

页      面：164-171页

核心收录：

学科分类：0710[理学-生物学] 0830[工学-环境科学与工程（可授工学、理学、农学学位）] 07[理学] 08[工学] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种] 

基　　金：Natural Science Foundation of Beijing Municipality: 5030001 Major State Basic Research Development Program of China: G20000161 

主　　题：transgenic cloning, GFP, Pig, early development. 

摘      要：In the present study, nuclear transferred embryos (NTEs) were reconstructed by using pig fetal fibroblasts as donors; in vitro matured oocytes as recipients. The effects of G418 selection on donor cells, duration of IVM of prepubertal gilt oocytes; oxygen tension in IVM of oocytes were investigated. The results were as follows: (i) When G418 selected cells expressing GFP were used as donors, the cleavage rate of NTEs decreased drastically in comparison to NTEs derived from donors without antibiotic selection (47.5% vs. 71.6%, p0.05). For the blastocyst rate, no significant difference was observed between two groups (10% vs. 10.4%, p0.05). However, no statistical difference was observed between NTEs derived from oocytes of 36 h IVM group; NTEs from oocytes of 42 h IVM group in the rates of cleavage (59.3% vs. 73.6%, p0.05); blastocyst formation (9.3% vs. 13.2%, p0.05); (iii) no significant difference was observed between NTEs reconstructed from oocytes matured under lower oxygen (7% O2) tension; NTEs derived from oocytes matured under higher oxygen tension (20% O2) in cleavage rate (70.6% vs. 67.1%, p0.05); blastocyst rate (11.8% vs. 12.3%, p0.05). These results suggest that: (i) G418 selection does not have a significant effect on cleavage rate of NTEs expressing GFP. (ii) Nuclear maturation is greatly improved by prolonging IVM duration from 36 to 42 h, while no significant differences were observed for developmental potential of transgenic embryos. Thus IVM 42 h is the better choice in order to obtain maximum number of MII oocytes as recipients. (iii) Lower oxygen tension; higher oxygen tension in IVM have no significant effect on development of cloned embryos.