Mao Chun-Ming, Yang Xiao, Cheng Xuan, Lü Ya-Xin, Zhou Jiang, Huang Cui-Fen
Genetic Laboratory of Development and Diseases, Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
Yi Chuan Xue Bao. 2003 May;30(5):407-13.
A keratinocyte-specific Cre transgenic construct (pK5-Cre) containing the keratin 5 promoter, Cre recombinase gene and polyA of human growth hormone gene was generated. The 4.2 kb DNA fragment of K5-Cre-hGH was introduced into 720 fertilized zygotes by microinjection. 695 injected eggs were implanted into the oviduct of 29 female mice respectively, from which 48 off-spring were obtained. Twelve mice carrying the transgene were identified by genotyping, and the integration efficiency is 25%. The K5-Cre transgenic mice were crossed with Smad4 conditional gene targeting mice to check the tissue-specific expression of the Cre recombinase and the Cre mediated recombination in multiple tissues. The results showed that the Cre recombinase was expressed in skin tissue only and successfully mediated the recombination between the loxP sites in vivo.
构建了一种角质形成细胞特异性Cre转基因载体（pK5-Cre），其包含角蛋白5启动子、Cre重组酶基因和人生长激素基因的polyA。通过显微注射将4.2 kb的K5-Cre-hGH DNA片段导入720枚受精卵中。将695枚注射后的卵分别植入29只雌性小鼠的输卵管中，从中获得了48只后代。通过基因分型鉴定出12只携带转基因的小鼠，整合效率为25%。将K5-Cre转基因小鼠与Smad4条件基因打靶小鼠杂交，以检测Cre重组酶在多种组织中的组织特异性表达以及Cre介导的体内重组。结果表明，Cre重组酶仅在皮肤组织中表达，并在体内成功介导了loxP位点之间的重组。
