摘要
构建了含有软骨组织特异性Ⅱ型胶原A1启动子和Cre重组酶基因的转基因载体pcol2A1 Cre。 32 3枚小鼠受精卵经显微注射引入转基因片段后 ,分别移植至 14只假孕母鼠的输卵管使其发育。共得到仔鼠 5 2只 ,PCR鉴定结果显示其中 10只小鼠基因组上有Cre基因的整合 ,整合率为 19 2 %。用整合有Cre基因的转基因小鼠与基因组上携带LoxP位点的条件基因打靶小鼠交配 ,以检测Cre酶介导的重组及其组织特异性。PCR结果表明 :col2A1 Cre转基因小鼠软骨组织中表达的Cre重组酶成功地介导了LoxP之间的重组。
A chondrocyte specific transgenic construct (pcol2A1 Cre) containing the cartilage specific typeⅡ collagen A1 promoter, the Cre recombinase gene and the polyA of human growth factor gene was generated. The 9 3kb DNA fragments were recovered from Not Ⅰ digested fragments and microinjected into 323 fertilized eggs. The injected eggs were implanted into the oviduct of 14 female mice. In the 52 offsprings, there were 10 mice carring the transgene identified by PCR, and the efficiency was 19 2%. The col2A1 Cre transgenic mice were crossed with a conditional gene targeting mice to check the Cre mediated recombination in multiple tissues. The results of the PCR analysis suggested that the Cre recombinase was expressed only in cartilaginous tissue and could mediate the recombination between the LoxP sites in vivo. The result was further confirmed by Southern blot.
基金
国家自然科学基金 (3 0 0 70 83 7)
国家杰出青年科学基金 (3 0 0 2 5 0 2 8)
国家高技术研究发展计划 (10 2 0 8 0 8 0 2 ) ~~
