[Construction of the oocyte-specific expressing phiC31 integrase vector pZP3-INT and its expression in mouse oocytes].
Streptomyces phage phiC31 integrase is a site-specific recombinase, which can catalyze site-specific, unidirectional recombination between the attP site and attB site. To explore whether it can be used to mediate the recombination of specific gene in oocytes, GV-stage oocytes were collected from 3-week-old Kunming White mice by puncturing antral follocles with a sharp needle, and micro-injected with oocyte-specific expressing phiC31 integrase vector pZP3-INT and site -specific recombination detection vector pBCPB+. phiC31 integrase mRNA were detected by RT-PCR and the recombination of pBCPB+ was evaluated by PCR in mouse oocytes at 48 h after injection. Both can get corresponding bands. These results indicated that the expression of phiC31 integrase can be driven by ZP3 promoter efficiently and phiC31 integrase can mediate the site-specific recombination between attP site and attB site in mouse GV-stage oocytes. It could be a powerful tool for the study of recombination of specific gene in mouse oocytes and would provide an alternative way for the mouse oocyte genome manipulation.
Xu
Huan-Yu
HY
Shanghai Institute of Medical Genetics, Shanghai JiaoTong University, Key Laboratory of Embryo Molecular Biology, Ministry of Health, Shanghai 200040, China. xuhuanyu@126.com
Gong
Xiu-Li
XL
Guo
Xin-Bing
XB
Ma
Qing-Wen
QW
Zeng
Yi-Tao
YT
D000818:Animals; D001435:Bacteriophages / Q000235:genetics; D001665:Binding Sites; D004527:Egg Proteins; D015870:Gene Expression; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D019426:Integrases / Q000235:genetics* / Q000378:metabolism; D008562:Membrane Glycoproteins; D051379:Mice; D009865:Oocytes / Q000201:enzymology / Q000378:metabolism / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D011956:Receptors, Cell Surface; D000072216:Zona Pellucida Glycoproteins
D004527:Egg Proteins; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D000072216:Zona Pellucida Glycoproteins; C000606521:Zp3 protein, mouse; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D019426:Integrases