关键词: Vill基因, pEF6/V5His-Vill, 显微注射, 转基因小鼠

Abstract: Abstract：In order to generate Vill transgenic mouse for the functional study of Vill gene, the 2 605bp total length of Vill gene was amplified by RT-PCR and cloned into the pMD19-T vector to construct pMD19-T-Vill vector at first. The target Vill fragment with proper restriction endonuclease sites was obtained by PCR amplification using specially designed primers and pMD19-T-Vill template. This target Vill fragment and pEF6/ V5-His vector were digested respectively by SpeI and BstBI in the same time,and then were connected together to construct pEF6/ V5-His-Vill expression vector. After target gene was verified to express in L929 cells, 4.5 kb length of transgenic construction including hEF-1α promoter、Plcd1、V5、His-tag and BGH was isolated and microinjected into 390 zygotes to produce77 offspring. Finally,16 hereditable lines were established from 19 positive transgenic founder mice. General appearance was not found difference between Vill-transgenic mice and the wild type mice. The Vill transgenic mouse is potential material for the study of Vill gene functions.

Key words: Vill gene, pEF6V5His-Vill, microinjection, transgenic mouse